Objective To study the possibility of salivary microbiota model to diagnose laryngopharyngeal re-flux(LPR).Methods A case-control study was applied to enroll 34 patients as case group who showed significant efficacy after 8 weeks of proton pump inhibitor treatment from February 2022 to November 2022.And 47 healthy volunteers matched by age,gender and body mass index with the case group were enrolled as the control group.Their salivary samples were collected before medication,and the salivary microbiota was detected by 16S rDNA se-quencing.Bioinformatics analysis was conducted on the sequencing results to compare species differences at the ge-nus level.A total of 24 patients and 33 cases in the control group were selected as train set and the rest as test set.Random forest method was used to classify data and ten fold cross validation was applied to select the optimal bacte-rial genus combination to construct a diagnostic model.The probability of disease(POD)index was calculated and receiver operating characteristic curve(ROC)was used to evaluate the diagnostic model in diagnosis of LPR.SPSS 18.0 software was utilized for statistical analysis.Results Compared with the control group,there was a statistical difference in the relative abundance of 22 genera in saliva between the case group and the control group(P＜0.05).A diagnostic model consisting of 6 genera was constructed,namely Lactobacillus,Novosphingobium,Bacillus,Pseudoalteromonas,Ralstonia and Phocaeicola.The area under the ROC curve of the test set was 0.843,the sensi-tivity of the diagnostic model was 60.0％,the specificity was 87.71％,and the Kappa value was 0.470.Conclusion The bacterial combination diagnostic model constructed from saliva microbiota based on microbiome and machine learning can effectively distinguish LPR patients from healthy individuals,which has potential clinical application value.

In order to prepare monoclonal antibodies with blocking activity against feline TNF-α,this study successfully constructed,expressed and purified the recombinant plasmid pET-28a-sTNFα based on the soluble feline TNF-α(sTNFα)gene,and further investigated the induced ex-pression.The conditions were explored and optimized to identify its biological activity;secondly,the feline TNF-α recombinant protein was used as an immunogen for mouse immunization,after cell fusion,screening of blocking active hybridoma cells and ascites preparation,the obtained mon-oclonal antibodies were tested.The results showed that the pET-28a-sTNFα plasmid was success-fully constructed and the bioactive feline TNF-α recombinant protein was expressed in E.coli sys-tem.The molecular weight was 34 kDa and the 50％inhibitory concentration was 1.22 pg/L.Three monoclonal antibodies(A6-B7-9,H5-E2-94 and C8-A10-100)with blocking activity were success-fully screened out.The results of Western blot showed that all the three mAbs could specifically bind to TNF-α with a titer of 1:512 000.When the concentration of the three mAbs was 100 mg/L,the inhibitory effect on TNF-α was the strongest.In this study,we screened antibodies that can block the activity of cat TNF-α,in order to provide novel,safe and effective candidate drugs for the treatment of TNF-α mediated diseases in cats.

BackgroundSuicide is one of the serious public health problems worldwide. The relationship between suicide and neutrophil-to-lymphocyte ratio （NLR） may vary in different regions and different age. It is necessary to further investigate the relationship between NLR and suicidal ideation in Chinese children and adolescents with depression. ObjectiveTo explore the correlation between NLR and suicidal ideation in children and adolescents with depression， so as to provide clues for exploring the biomarkers of suicide. MethodsA retrospective analysis of 536 children and adolescents with depression who were hospitalized in the Third People's Hospital of Fuyang from January 2020 to December 2022 and met the diagnostic criteria of the International Classification of Diseases， tenth edition （ICD-10） was performed. Patients were divided into two groups according to whether they reported suicidal ideation. Demographic data， discharge diagnosis， Hamilton Depression Scale-17 item （HAMD-17） score and hematological test data （neutrophil counts， lymphocyte counts） on the second day were collected from medical records. Receiver operating characteristic （ROC） curve was used to determine the optimal cut-off point of NLR for predicting suicidal ideation in children and adolescents with depression， and binary Logistic regression was used to analyze the risk factors for suicidal ideation. ResultsAmong the 536 patients， 429 cases （80.04%） had no suicidal ideation， and 107 cases （19.96%） had suicidal ideation. Compared with patients without suicidal ideation， the HAMD-17 score ［（25.28±8.86） vs. （21.21±8.46）， F=19.400， P<0.01］， neutrophil level ［（3.85±1.68）×10⁹/L vs. （3.15±1.14）×10⁹/L， Z=4.073， P<0.01］， and NLR level ［（1.96±1.50） vs. （1.52±0.71）， Z=3.532， P<0.01］ in the suicidal ideation patients were significantly higher. The optimal critical NLR value determined by the ROC curve was 1.52 （59.80% sensitivity， 58.50% specificity）， with an area under the curve of 0.610. Logistic regression analysis showed that the risk of suicidal ideation was 1.94 times higher in those with high NLR than in the low NLR after controlling for age， sex， age at onset， duration of illness， and HAMD-17 score （OR=1.940， 95% CI： 1.251~3.009， P=0.003）. ConclusionNLR may be a risk factor and potential biomarker influencing suicidal ideation in the children and adolescents with first-episode depression. ［Funded by Scientific Research Project of Fuyang Municipal Health Commission （number， FY2020xg14）］

Objective: To investigate knowledge, attitudes, and practice on exercise and dietary behaviors among the primary and middle school pupils in Shandong province, and to provide a theoretical basis for a targeted education on diet and physical health among the primary and middle school students. Methods: A questionnaire survey was conducted among 6 161 students from 3 cities in Shandong province by random cluster sampling. Results: Self-reported rate of having breakfast every day for primary and middle school students were 85.0% and 75.3% ,respectively(χ2=84.0，P<0.01). 41.2% of primary school students were reported to do physical exercises more than 60 minutes every day and the rate was only 29.6% for middle school students(χ2=270.0，P<0.01). 39.2% of boys were reported to do physical exercises more than 60 minutes every day and the rate was 28.7% for the girls(χ2=90.4，P<0.01). The awareness rates of knowledge about dietary and physical exercise among primary and middle school students were 44.4% and 55.8%, respectively(χ2=702.0，P<0.01). The awareness rates among boys and girls were 50.4% and 51.9% respectively (χ2=12.4，P<0.01). Parents’ occupation, knowledge score and education level of mother were influencing factors of breakfast behavior everyday. The physical exercise was influenced by gender, parents' occupation, knowledge score and education level of mother. Conclusion It is necessary to educate students to take healthy diet and have more physia1 activities. Families, schools and government departments need to take feasible measures to ensure the time and intensity of pupils’ physical exercise through many ways.

Objective To establish the method for quality control of ethyl-acetate parts of Ferula sinkiangesis. Methods HPLC was used to detect the contents of ferulic acid, farnesiferol A and farnesiferol C. Waters XTerra RPC18 column (250 mm×4.6 mm, 5 μm) was used; acetonitrile-0.1% phosphoric acid was as mobile phase with gradient elution; flow rate was 1.0 mL/min; detective wavelength was 324 nm; temperature was 30℃; sample volume was 10 μL ResultsFerulic acid, farnesiferol A, and farnesiferol C showed a good linear relationship range from 0.05– 1.0 mg/mL, 0.132–2.64 mg/mL, and 0.118–2.36 mg/mL, respectively. The average recovery rates were 99.34%, 98.96% and 99.24% respectively. The contents of ferulic acid, farnesiferol A and farnesiferol C were 33.4, 76.5, 72.3 mg/g respectively.Conclusion The method was simple, accurate and repeatable, with good repeatability and stability, which can be used for the quality control of ethyl-acetate parts ofFerula sinkiangesis.

Objective To analyze the correlation of tumor recurrence after liver transplantation for hepatocellular carcinoma (HCC)with the expression levels of regulatory T cell (Treg)and cytokines in peripheral blood. Methods A total of 56 patients who underwent liver transplantation in the 309th Hospital of People's Liberation Army from 2010 to 2014 were studied. According to the postoperative pathological data,all the patients were divided into the group of liver transplantation for HCC (HCC group,n=28)and group of liver transplantation for cirrhosis (liver cirrhosis group,n=28), of which the HCC group was further divided into non-recurrence group (n=8)and recurrence group (n=20)according to the situation of postoperative tumor recurrence. The expression levels of Treg and cytokines [vascular endothelial growth factor (VEGF),interleukin (IL)-2,IL-10,IL-12,transformation growth factor (TGF)-βand interferon (IFN)-γ]in peripheral blood of the patients in various groups were compared. Results Compared with the liver cirrhosis group,levels of IFN-γand IL-12 in the non-recurrence group increased significantly (both P<0.05);levels of Treg%,VEGF,IFN-γ, IL-10 and TGF-βin the recurrence group increased significantly,while levels of IL-2 and IL-12 decreased significantly (all P<0.05). Compared with the non-recurrence group,levels of Treg%,VEGF,IL-10 and TGF-βin the recurrence group increased significantly,while levels of IFN-γ,IL-2 and IL-12 decreased significantly (all P<0.05 ). Conclusions Levels of Treg and cytokines can be used to predict the tumor recurrence after liver transplantation for HCC.

Objective To explore the clinical manifestations,diagnosed main points,laparoscopic trestment strategy of ectopic appendicitis.Methods The 19 clinical data of ectopic appendicitis,from April 2011 to April 2015 in our hosipital,were retrospective analysised,and we sum up the clinical manifestation,diagnosis and treatment,laparoscopic exploration indications and the technique of laparoscopic appendectomy.Laparoscopy exploration and laparoscopic appendectomy were made on all patients under general anesthesia by tracheal cannula.Results Of 19 patients,16 cases by ultrasonic examination get correct diagnosis before operation,of which the ultrasound diagnosis rate is 92.63％,3 limitations of acute peritonitis patients were preoperative misdiagno~s,they were the right kidney stones,acute cholecystitis and the right side of the annex inflammation;there were 2 patients converted to open operation,and the transfer rate is 10.53％.The blood loss of the operation is about 5-20 mL.They were ambulant after 9 hours,in full flow diet after 12 hours,and the average hospitalization time is 4 days.One patient,no other long-term complications were found postoperatively follow-up of 12 to 18 months,were poke holes in infection,and the complications of laparoscopic surgery is 5.27％.Conclusions Ectopic acute appendicitis has no specific clinical manifestation,and is easy to misdiagnosis.In the auxiliary examination,ultrasound is the first choice for the diagnosis of ectopic appendictis.Laparoscopy exploration and laparoscopic appendectomy are the ideal treatments of emergency treatment of ectopic appendicitis,and are worth of clinic application.

Objective To evaluate the efficacy and security of splenic artery ligation for severe hypersplenism during liver transplantation.Method Thirty-two liver transplant patients with preoperative hypersplenism were selected,including 17 cases (ligation group) treated by splenic artery ligation during liver transplantation,and rest 15 patients as non-ligation group.The fluctuation of white blood cells,platelets and volume of spleen were compared between these two groups.At the same time,splenic infarction,postoperative infection,recurrent gastrointestinal bleeding,splenic artery steal syndrome and other complications were observed in these two groups.Result All recipients were followed up for over 6 months.One recipient in ligation group died of multiple organ dysfunction caused by delayed recovery of liver donor with the survival rate being 94.1％ (16/17).The survival rate in non-ligation group was 93.3 ％ (14/15) (one recipient died of respiratory failure caused by pulmonary infection).There was no statistically significant difference in survival rate between these two groups (P＞0.05).Splenic necrosis wasn't detected in the ligation group.The splenic volume in ligation group was significantly less than that in non-ligation group (P＜0.01).The products of splenic maximum length and wide diameter shrunk 33.17-± 8.26 cm2 and 22.47 ± 7.25 cm2 in ligation group and non-ligation group,respectively.The platelet counts of ligation group were significantly greater than those of non-ligation group in all the observation points within 6 postoperative months (P＜0.01).The white blood cell counts of ligation group were greater than those of non-ligation group at the first week postoperatively (P＜0.01),whereas,there was no statistically significant difference between these two groups from then on (P＞0.05).The infection incidence of ligation group was lower than that of non-ligation group within 6 postoperative months (P ＜0.05).Statistically significant differences in recurrent gastrointestinal bleeding and splenic artery steal syndrome weren't found between these two groups (P＞0.05).Conclusion Splenic artery ligation in liver transplantation is safe and effective.It can rapidly increase the counts of platelet and white blood cell in the earlier postoperative time,which is beneficial to patient's recovery.Besides,it adds no correlative complication.

Objective To establish a high-throughput detection of medicine on DPPH free radical scavenging activity;To analyze and preliminarily screen the anti-oxidant activity part of Ferula sinkiangensis. Methods Taking DPPH free radical scavenging rate of ascorbic acid as positive control, and IC50 as the evaluating criterion of DPPH free radical scavenging capability, oxidation resistance of different polar parts (petroleum ether part, ethyl acetate part, methyl alcohol part, and water part) of Ferula sinkiangensis was evaluated and screened. Results Different polarity parts of Ferula sinkiangensis DPPH radical scavenging of IC50 were:petroleum ether part 3252.22 μg/mL, ethyl acetate part 36.22 μg/mL, methyl alcohol part 32.22 μg/mL, water part 2643.38 μg/mL, and positive control group ascorbic acid 27.16 μg/mL. Conclusion The ethyl acetate and methyl alcohol parts of Ferula sinkiangensis were effective anti-oxidant activity site to eliminate DPPH free radicals. In this study, a simple, sensitive, and high-throughput detection method of DPPH radical scavenging assay was established to provide reference for the anti-oxidation medicine detection screening.