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 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 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 explore the efficacy and safety of thymalfasin in the treatment of severe pulmonary infection after liver transplantation.Methods Twenty seven patients who developed severe lung infection after undergoing liver transplantation in Organ Transplant Institute of the 309 th Hospital of People’s Liberation Army from January 2008 to May 2014 were enrolled in this study.According to whether the application of thymalfasin,the patients were divide into thymalfasin group (n =11)and control group (n =16).In the thymalfasin group,thymalfasin was administered via subcutaneous injection at a dose of 1.6 mg once daily for consecutive two weeks.In the control group,conventional anti-infection therapy was delivered. Ventilator time,duration of fever,the length of intensive care unit (ICU)stay and mortality were statistically compared between two groups.And the incidence of acute rejection (AR)was monitored.Results Ventilator time,duration of fever,length of ICU stay of patients in the thymalfasin group were significantly shortened compared with those in the control group (all in P <0.05).There was no significant difference in the mortality between two groups.No clinical AR was observed in either group.No thymalfasin-related adverse event was found in the thymalfasin group.Conclusions Thymalfasin can improve the curative effect to anti-infection of patients with severe pulmonary infection after liver transplantation without the incidence of AR,which is efficacious and safe in the treatment of severe pulmonary infection.

Objective To discuss the relationship between Foxp3 +regulatory T cell (Treg)and tumor recurrence of patients after liver transplantation for primary hepatocellular carcinoma (HCC) over University of California,San Francisco (UCSF)criteria.Methods Clinical data of 24 patients with HCC undergoing liver transplantation in the Organ Transplantation Research Institute of the 309 th Hospital of People's Liberation Army from January 2010 to December 2013 were retrospectively studied.During the follow-up,4 patients recurred (tumor recurrence group)and other 20 patients did not recur (tumor non-recurrence group).The blood samples of healthy people was selected as control group at the same period.The levels of alpha-fetoprotein (AFP)were compared at different time points of the recurrence group and the non-recurrent group before and after transplantation.The levels of Foxp3 +Treg (Foxp3 +Treg%)were compared at different time points of the tumor recurrence group,the tumor non-recurrence group and the control group before and after transplantation.The relations between expression of Foxp3 +Treg and the levels of AFP,CD3 + and CD8 +T before and after transplantation were analyzed by correlation analysis.Results Compared with the level of Foxp3 +Treg before transplantation and the normal level,the expression of Foxp3 +Treg of patients in tumor non-recurrence group after transplantation firstly decreased,then gradually increased and finally stabilized at a low level.Compared with patients in tumor non-recurrence group,the levels of AFP and Foxp3 +Treg of patients in tumor recurrence group increased obviously and were significantly higher than the normal levels (both in P <0.01).Moreover,abnormal increase of Foxp3 +Treg at early stage was prior to AFP among the patients in tumor recurrence group.Correlation analysis indicated that the change of Foxp3 +Treg was consistent with the changes of AFP,which was positively correlated (P <0.01).But the change of Foxp3 +Treg was contrary to the change of effector T cells (CD3 +T cells and CD8 + T cells),which was negatively correlated (P <0.05-0.01).Conclusions Foxp3 +Treg is closely associated with tumor recurrence after liver transplantation for HCC.In the patients after liver transplantation for HCC over UCSF criteria,the higher Foxp3 +Treg is,the higher the recurrence risk is.Joint detection of AFP is beneficial to find tumor recurrence.

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.

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.