AIM:To develop a HPLC method of determining osthole and imperatorin in Compound Shajiziyou Suppository(oil of Semen Hippophae,Fructus Cnidii,Radix Sophorae flavescentis,etc.). METHODS: Phenomnex(luna) C_(18) column was used at(25 ?C).The mobile phase consisted of methanol-water(65(∶)35).The flow rate was 1.0 mL/min.The detection wavelength was set at 310 nm. RESULTS: The linear ranges of osthole and imperatorin were 0.051-0.816 ?g(r=0.999 6) and 0.026-0.416 ?g(r=0.999 8),respectively,The average recoveries were 98,4% with RSD of 1.4% and 97.6% with RSD of 2.0% respectively. CONCLUSION: This method is simple,accurate,reproducible and can be used for the determination of osthole and imperatorin in Compound Shajiziyou Suppository.

Objective:To investigate the anticaries effects of hesperidin in rats.Methods:The MIC of hesperidin against Streptococ-cus sobrinus(S.sobrinus)was explored with disc diffusion method.The rats with artificial caries were administered with 1/2 MIC hes-peridin,0.12%Chlorhexidine and distilled water respectively.The results of S.sobrinus level,Keyes scoring and DIAGNOdent exam-ination were used to evaluate the effects of hesperidin on S.sobrinus and caries development.Results:The MIC of hesperidin agaist S. sobrinus was 8 mg/ml.The S.sobrinus level was not statistically different between hesperidin group and negative control group(P>0. 05).Keyes scores of grade E of smooth and pit and fissure caries in hesperidin group were lower than in negative control(P<0.05), those of grade Ds and Dm of pit and fissure caries in hesperidin group were lower than in negative control(P<0.05 ),while higher than in chlorhexidine group(P<0.05).Hesperidin showed significant anti-caries effect(P<0.05)examined by DIAGNOdent.Con-clusion:Hesperidin at 1/2 MIC has anti-caries effect in rats without influence of the oral microecological balance.

Objective: To analyze the factors affecting frailty among convalescent elderly population, so as to provide the evidence for prevention and treatment of frailty. Methods: The convalescent elderly population at ages of 60 years and older were selected from the Wuyunshan Hospital in Hangzhou City using the convenience sampling method from April 2022 to April 2024. Demographic information, chronic disease and medication were collected using questionnaire survey. Frailty was assessed using the FRAIL Scale. Factors affecting frailty among convalescent elderly population were analyzed using a multivariable ordinal logistic regression model. Results: A total of 1 050 questionnaires were distributed, and 1 023 valid questionnaires were collected, with a response rate of 97.43%. There were 793 males (77.52%) and 230 females (22.48%); 192 respondents aged 60 to <70 years (18.77%), 431 respondents aged 70 to <80 years (42.13%) and 400 respondents aged ≥80 years (39.10%); 718 respondents with university degree (70.19%); 890 respondents with a monthly income of 10 000 yuan to <20 000 yuan (87.00%); 130 respondents with comorbidity and polypharmacy (12.71%); and 197 respondents with the risk of malnutrition (19.26%). There were 202 cases with pre-frailty (19.75%) and 47 cases with frailty (4.59%). Multivariable ordinal logistic regression analysis showed that the convalescent elderly population who were aged ≥80 years (OR=3.710, 95%CI: 2.340-5.883), with comorbidity and polypharmacy (OR=12.370, 95%CI: 7.949-20.369) and with the risk of malnutrition (OR=5.414, 95%CI: 3.691-7.933) had higher risk of frailty. Conclusion The high risk of frailty among convalescent elderly population is associated with age, comorbidity and polypharmacy, and malnutrition.

Objective To investigate the occurrence of hepatocyte autophagy and the expression of autophagy associated proteins at different time points in sepsis mice model established by cecal ligation and puncture.Methods Fifty male ICR mice aged 6-8 weeks were randomly (random number) divided into 5 groups:CLP 3 h group,CLP 6 h group,CLP 12 h group,CLP 24 h group and control group.The samples were collected at the corresponding time points after operation.Liver function was tested to obtain alanine aminotransferase (ALT) and aspartate aminotransferase (AST).The expression of TNF-α and IL-6 was detected by ELISA,the inflammation of liver tissue by HE staining while the ultrastructure of autophagosome and autolysosome in liver tissue was observed by electron microscopy.Meanwhile,the expression of LC3 and P62 was detected by Western blot.and the location and quantity of autophagosome was observed by Immunofluorescence.SPSS 21.0 was used for statistical analysis.Two independent sample t tests were performed to compare the two groups,and one-way ANOVA was used for comparison between multiple groups.Results ALT and AST increased significantly after CLP,and peaked at 12 h,respectively (137.8 ±11.94) U/L and (475.3 ±57.34) U/L.The expression of IL-6 peaked at 6 h (2589.63 ±27.96) pg/mL,and TNF-α peaked at 3 h (320.21±8.9) pg/mL.HE staining showed activation of Kupffer cells and infiltration of single granulocyte.Electron microscope showed the formation of autophagosome and autolysosome.Compared with the control group,the ratio of LC3 Ⅱ/Ⅰ increased first and then decreased,peaked at 6 h (t=13.35,P＜0.05).The content of P62 protein declined first and then went up,reaching the lowest level at 6 h (t=66.1 l,P＜0.05),contrary to the trend of LC3 Ⅱ/Ⅰ.Immunofluorescence results illustrated that the fluorescence intensity of LC3 reached its high point at 6 h (t=12.52,P＜0.05).Conclusions Autophagy occurred in the liver of sepsis mice model established by cecal ligation and puncture.The formation of autophagosome first increased and then decreased,reaching its peak at 6 h.

OBJECTIVE To investigate whether quinolinic acid(QA)induces autophagy in PC12 cells and its relationship with glycogen synthase kinase-3β(GSK-3β)/β-catenin related signaling path?ways. METHODS PC12 cells were treated with QA 2.5,5.0 and 10.0 mmol·L-1 for 24 h. The cell viability was determined by MTT assay. Autophagy fluorescent spots labelled form of microtubule-associated protein 1 light chain 3(LC3)was examined by LC3 immunostaining. The expressions of GSK-3β,β-catenin,LC3 and Beclin 1 were determined by Western blotting. RESULTS QA inhibited PC12 cell survival in a concentration-dependent manner,and IC50 was 8.7 mmol · L- 1. Compared with normal control group,QA 2.5,5.0 and 10.0 mmol · L-1 increased autophagic intracellular LC3 fluorescence spots,elevated the expression ratio of LC3-Ⅱ/LC3-Ⅰ and expression of Beclin 1 in PC12 cells(P<0.05). In addition,QA enhanced GSK-3βexpression and decreasedβ-catenin expression(P<0.05,P<0.01). CONCLUSION QA induces autophagy in PC12 cells. This mechanism may be associated with the activation of GSK-3β/β-catenin related signaling pathways.

Objective To investigate the effect of fetoscopic photocoagulation of communicating placental vessels in twin-twin transfusion syndrome(TTTS)(selective or non-selective) on the perinatal outcomes.Methods Six cases of TTTS admitted in our department from Dec.2006 to Jun.2008 underwent fetoscopic photocoagulation of communicating vessels.Under direct real-time sonographic guidance,a 3-mm-diameter fetoscope was percutaneously inserted through the maternal abdominal wall into the amniotic cavity of the recipient twin.A combination of ultrasonographic and fetoscopic vision was used to identify the crossing vessels which were systematically coagulated using Nd:YAG laser fiber or bipolar electrocoagulation.Results All the 6 mothers tolerated the procedure without major complications.Two fetal survival rate was 33.33%.Conclusion Fetoscopic photocoagulation of communicating placental vessels in TTTS can effectively improve perinatal outcomes.

Objective:To study the influence of serum triggering receptor expressed on myeloid cells 1(TREM-1)level on prognosis in elderly patients with sepsis and acute respiratory distress syndrome(ARDS).Methods:A total of 100 elderly patients with sepsis were selected as the research objects.All the patients with sepsis were divided into sepsis ARDS group and sepsis non-ARDS group.General data and TREM-1 level were compared between the two groups.The patients with sepsis ARDS were divided into death group and survival group according to the survival status during the 28-day follow-up.TREM-1 level, acute physiology and chronic health evaluation(APACHE)Ⅱ score and SOFA score were compared between the two groups.The correlation between serum TREM-1 level and procalcitonin(PCT), APACHE Ⅱ score and SOFA score was analyzed.The survival rate of high TREM-1 level group and low TREM-1 level group was compared.Results:The age, white blood cell(WBC), PCT, APACHE Ⅱ score, SOFA score and TREM-1 level of sepsis ARDS patients were significantly higher than those of non-ARDS patients( t=2.722, 6.088, 11.55, 6.889, 4.661, 6.122, all P<0.05). The incidence of sepsis ARDS patients with chronic obstructive pulmonary disease was significantly higher than that of non-ARDS patients( χ2=7.895, P<0.05). Serum TREM-1 level, APACHE Ⅱ score and SOFA score of ARDS patients in death group were significantly higher than those in survival group( t=3.293, 6.173, 4.255, all P<0.05). Serum TREM-1 level was positively correlated with PCT, APACHE Ⅱ score and SOFA score( t=0.553, 0.602, 0.636, P<0.001). The Kaplan-Meier survival curve showed that the survival rate of high TREM-1 level group was significantly lower than that of low TREM-1 level group( χ2=3.999, P=0.036). Cox regression analysis showed that TREM-1 level was a risk factor for the prognosis of ARDS patients with sepsis( HR=1.893, 95% CI: 1.049-3.414). Conclusions:Serum TREM-1 level is significantly increased in elderly patients with sepsis ARDS, which is closely related to the prognosis and can be used as a potential prognostic biomarker.

Objective To analyse the differential metabolites and related metabolic pathways in stable angina pectoris of coronary artery heart disease with spleen deficiency and phlegm turbidity syndrome by serum metabolomics.Methods This study observed 60 patients with stable angina pectoris of coronary artery heart disease with spleen deficiency and phlegm turbidity syndrome and 60 healthy volunteers in the same period.Liquid chromatography-mass spectrometry(LC-MS)was performed on the serum metabonomics.The differential metabolites were identified by multivariate statistical analysis of the original spectrogram and original data,and enrichment analysis of KEGG metabolic pathway was analyzed.Results A total of 60 patients in the group of stable angina pectoris of coronary artery heart disease with spleen deficiency and phlegm turbidity syndrome participated in the study,and a total of 60 healthy volunteers in the control group participated in the study.There was no statistical difference in general information and biochemical indicators between the two groups(P>0.05);Eighteen differential metabolites were found respectively,including phenylacetaldehyde,orthophosphate,guanosine,diethyl phosphate,2-dehydro-d-gluconate,guanine and 5-(2-hydroxyethyl)-4-methylthiazole down-regulated expression,taurocholate,2-propylglutaric acid,8-amino-7-oxononanoate,l-tyrosine,s-sulfo-l-cysteine,cyclohexanecarboxylic acid,porphobilinogen,(r)-acetoin,octanoylglucuronide,melatonin and solanine up-regulated expression,involving phenylalanine metabolism,thiamine metabolism,purine metabolism.Conclusion The differential metabolites reveal the metabolic essence of stable angina pectoris of coronary artery heart disease with spleen deficiency and phlegm turbidity syndrome from the micro level,and can provide clues for clinical early warning of patients with stable angina pectoris of coronary artery heart disease with spleen deficiency and phlegm turbidity syndromet.

Objective:To investigate the protective effect and mechanism of hydroxysafflor yellow A (HSYA) on severe acute pancreatitis (SAP) related lung injury.Methods:Fifty mice were randomly (random number) divided into five groups: the sham-operated group, SAP group and different doses (20, 40 and 80 mg/kg) of HSYA pretreatment group. Mice were pretreated with HSYA 24 h before SAP induction, pancreatic and lung tissues were isolated for histopathological examination at 72 h after modeling, and bronchoalveolar lavage fluid (BALF) was collected for biochemical analysis. Results:Compared with the sham-operated group, serum amylase activity, lung injury pathological score and BALF protein concentration in the SAP group were significantly increased [(2120.44 ± 354.50) U/L vs. (226.72 ± 20.84) U/L; (6.91 ± 0.28) vs. (0.53±0.18); (2563.25±348.22) μg/mL vs. (345.62±56.35) μg/mL, all P<0.05]. Inflammatory factors tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels and myeloperoxidase (MPO) activity were increased [(120.5±14.25) pg/mL vs. (31.5±4.82) pg/mL; (214.72±10.62) pg/mL vs. (39.26±5.66) pg/mL; (4.52±0.34) U/mg vs. (1.03±0.17) U/mg]. Compared with the SAP group, HSYA pretreatment significantly attenuated SAP-related pancreatic and lung tissue damage and the activities of the inflammatory factors TNF-α, IL-6 and MPO in BALF. In addition, HSYA promoted the expression of the antioxidant protein heme oxygenase-1 and blocked the activation of the NF-κB signaling pathway. Conclusions:HSYA exerts anti-inflammatory and antioxidant activities to inhibit SAP-related lung injury, which indicated that HSYA may be a potential therapeutic drug for SAP-induced lung injury.

Objective:To compare the early and late predictive value of several critical illness scores (CISs) and biomarkers in patients with bloodstream infection (BSI)-associated pneumonia, and to identify the value of procalcitonin (PCT) in etiological diagnosis.Methods:Patients with at least one positive blood culture within 24 hours admission to department of emergency of China-Japan Friendship Hospital from January 2014 to December 2018 and with final diagnosis of pneumonia were enrolled. Sequential organ failure assessment (SOFA), mortality in emergency department sepsis (MEDS), Logistic organ dysfunction system (LODS), and acute physiology and chronic health evaluationⅡ (APACHEⅡ) scores were calculated based on the first parameters on the day of admission. Differences of various indicators among different Gram-stained bacterial infections and among patients with different prognosis at 28-day or 60-day were compared. Receiver operating characteristic (ROC) curve was used to analyze the value of biomarkers in differential diagnosis of pneumonia caused by single bacterial infection, and the predictive value of several CISs and biomarkers on 28-day and 60-day death of patients with pneumonia.Results:Among 540 patients with pneumonia caused by single bacterial infection, 256 (47.4%) patients with Gram-positive bacteria (GPB) infection and 284 (52.6%) with Gram-negative bacteria (GNB) infection. The 28-day mortality was 29.4% (159/540) and the 60-day mortality was 36.3% (196/540). PCT level was significantly higher in patients with GNB infection than that in GPB infected patients [μg/L: 1.99 (0.32, 13.19) vs. 0.45 (0.13, 3.53), P < 0.01]. There were significant differences of CISs and biomarkers between death group and survival group in predicting 28-day and 60-day mortality in BSI-associated pneumonia. ROC curve analysis showed that: ① the optimal cut-off value of PCT in the diagnosis of single bacterial infection was 0.48 μg/L, with the area under ROC curve (AUC) was 0.739 [95% confidence interval (95% CI) was 0.686-0.793]. When PCT value was greater than 4.49 μg/L, the specificity of diagnostic of GNB infection could reach 81.8%, and the positive predictive value (PPV) was 75.0%. When PCT value was greater than 10.16 μg/L, the diagnostic specificity could reach 91.2%. ② In the prediction of 28-day and 60-day mortality, the SOFA score showed highest AUC [28-day: 0.818 (95% CI was 0.768-0.867), 60-day: 0.800 (95% CI was 0.751-0.849)]. SOFA score greater than 8.5 points could help to predict 28-day and 60-day mortality for pneumonia patients with specificity of 90.5% and 91.6%, respectively. AUC of PCT for predicting 28-day and 60-day mortality in patients with BSI associated with pneumonia was 0.637 (95% CI was 0.575-0.700) and 0.628 (95% CI was 0.569-0.688), respectively. When PCT value was greater than 8.15 μg/L, the specificity and negative predictive value (NPV) were 80.2% and 75.1% respectively, and they could reach 80.2% and 68.7% when PCT value was greater than 7.46 μg/L. Conclusion:PCT is more reliable in the identification of pathogen type in BSI-associated pneumonia, while CISs may be more advantageous in the assessment of early and late prognosis.