Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

Objective To observe the therapeutic effect and mechanism of external application with Shilian Powder combined with oral administration of Shumai Capsules for the treatment of rats with diabetic foot ulcer(DFU).Methods Eight male rats with successfully modeled foot ulcer(DF)were used as the control group.While 24 male rats with successfully modeled DFU were randomly divided into DFU group,Shumai Capsules group and Shilian Powder combined with Shumai Capsules group,with eight rats in each group.After the corresponding interventions,we determined the wound healing rate,histopathological changes of wound,levels of inflammatory factors such as interleukin(IL)-6,IL-1β and vascular endothelial growth factor(VEGF)in serum,levels of Apelin and Relaxin,and protein expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and VEGF in wound tissue,as well as mRNA expressions of PI3K,AKT,Relaxin and Apelin.Results Compared with the control group,the DFU group showed a significant decrease in wound healing rate,VEGF level in serum and wound,wound Relaxin level,protein and mRNA levels of wound AKT(P<0.05),and a significant increase in serum IL-6 and IL-1β levels,wound Apelin level,wound PI3K protein and mRNA levels(P<0.05),and the reduced wound granulation tissue and formation of new capillaries and increased inflammatory cell infiltration were seen under the microscope.Compared with the DFU group,the wound healing rate,VEGF level in serum and wound,wound Relaxin and Apelin levels,protein and mRNA levels of wound PI3K and AKT in the Shumai Capsules group and Shilian Powder combined with Shumai Capsules group were significantly increased(P<0.05),and serum IL-6 and IL-1β levels were significantly decreased(P<0.05),and the increased wound granulation tissue and formation of new capillaries and reduced inflammatory cell infiltration were seen under the microscope.Compared with the Shumai Capsules group,the wound healing rate,wound VEGF level,wound Relaxin and Apelin levels,protein and mRNA levels of wound PI3K and AKT in Shilian Powder combined with Shumai Capsules group were significantly increased(P<0.05),and the serum IL-6 and IL-1β levels were significantly decreased(P<0.05),and the increased wound granulation tissue and formation of new capillaries and reduced inflammatory cell infiltration were seen under the microscope.Conclusion External application with Shilian Powder combined with oral administration of Shumai Capsules can promote the wound healing in rats with DFU,its mechanism is related to the activation of PI3K/AKT/Relaxin/Apelin signaling pathway.

Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

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.

{L-End}Objective To explore the suitable methods for individual occupational stress examination and evaluation for workers in China based on the electronics industry. {L-End}Methods A total of 1 164 workers from four electronics enterprises in Beijing City were selected as the research subjects using a convenient sampling method. The Occupational Stress Measurement Scale, which was developed based on the Japanese Brief Job Stress Questionnaire, was used to assess the occupational stress of the research subjects, and test the reliability and validity of the scale. Percentile norms and T-score norms were established, and the T-scores of the three dimensions of stress reaction (psychological reaction and physical symptoms), stress factors, and social support were divided into five stages using the normal distribution method with x¯±0.5 s and x¯±1.5 s, which was used to explore a proposed standard for assessing individual stress levels. {L-End}Results The revised scale consisted of 57 items, which could explain 64.0% of the total variation of occupational stress based on the result of item analysis and exploratory factor analysis. The results of confirmatory factor analysis showed that the model fitted well, with root mean square error of approximation of 0.052, standardized root mean square residual of 0.070, comparative fit index of 0.960, and Tucker-Lewis index of 0.958. The Cronbach's α coefficient of the scale was 0.951, and the Spearman-Brown coefficient was 0.837. Cronbach's α coefficients of the three dimensions ranged from 0.794 to 0.952, and the Spearman-Brown coefficients ranged from 0.737 to 0.850. The scores of the three dimensions of stress reaction, stress factors, and social support were (49.1±13.0), (44.9±7.7), and (18.1±3.6), respectively. Workers in the electronics industry met one of the following items were identified as the high level of occupational stress individuals: i) a score of stress reaction dimension ≥72.0 points; ii) a sum of stress factor and social support dimension scores ≥81.0 points, and a stress reaction dimension score ≥58.0 points. {L-End}Conclusion The Occupational Stress Measurement Scale and the criteria for determining high level of occupational stress can be used to assess the individual occupational stress levels of workers in the electronics industry in China. This method can provide a theoretical basis for the formulation of occupational stress examination and assessment for Chinese workers.

The ways of treating diabetes mellitus with depression by using Traditional Chinese Medicine (TCM) include internal treatment, external treatment and combining internal and externalmethod, mostly based on the viscera syndrome differentiation and Qi blood phlegm stasis syndrome differentiation, focusing on soothing the liver qi and regulating, liver and spleen. The internal treatments mainly include self-made prescriptions or classic prescriptions, such as Jiawei-Xiaoyao Powder, Danggui-Shaoyao Decoction, Wendan Decoction, etc; External treatments mainly include common acupuncture, ear acupuncture, scalp acupuncture, abdominal acupuncture, acupoint application, etc. The above treatments can improve the symptoms of depression, and control the blood glucose level.

Objective:To explore the effect of Olmesartan on the antigen presenting function of dendritic cells (DCs) in rats.Methods:Bone marrow-derived dendritic cells of female Lewis rats were obtained. Bone marrow-derived dendritic cells were cultured with Olmesartan (final concentration 10 μmol/L; OLM-DCs) or equal volume of DMSO (Con-DCs). Mean fluorescence intensity (MFI) of the surface costimulatory molecule CD80, CD86 and MHCⅡ on DCs and the levels of IL-10 and TGF-β of DCs were analyzed by flow cytometry. DCs and CD4 +T lymphocytes were cocultured. T lymphocytes proliferation was analyzed by flow cytometry. IFN-γ in the supernatants were determined by ELISA. Results:The expression of MHC Ⅱ on DCs was inhibited by Olmesartan. The level of intracellular IL-10 in DCs was up-regulated by Olmesartan. Compared with Con-DCs, T lymphocytes proliferation and the level of IFN-γ were inhibited by OLM-DCs.Conclusions:Olmesartan could induce tolerogenic DCs in vitro. These tolerogenic DCs could inhibit T lymphocytes proliferation and the production of Th1 cytokine.

Objective To prepare recombinant plasminogen activator(Pla) protein in E.coli BL21 cells that can be used in studying interactions between Yersinia pestis proteins and immunologic diagnosis of plague.Methods The pla gene was amplified by PCR and cloned into the pET28a expression vector.E.coli BL21 competent cells were transformed with the recombinant vectors, and isopropyl-β-D-thiogalactopyranoside ( IPTG) was added to induce expression of Pla protein. The expressed protein was detected by SDS-PAGE electrophoresis.The inclusion bodies of Pla protein were denatured in 8 mol/L urea, and then refolded using gradient urea solutions.The purified protein was identified by SDS-PAGE electrophoresis and Western blot.Results and Conclusion The constructed expression vector was demonstrated to be correct through agarose gel electrophoresis and sequencing.The recombinant Pla protein was accumulated as an inclusion body in E.coli, and the overexpression product was mainly a target protein, the yield of which was very high.SDS-PAGE purity of the bioactive Pla protein was obtained by denaturing and refolding the inclusion bodies.This study provides a simple and quick method for highly efficient preparation of biologically active Pla protein.

Objective To establish a simple and quick loop-mediated isothermal amplification (LAMP)method for detection of Yersinia pestis.Methods LAMP Primers were designed based on the specific sequence 3a in Y.pestis chromosome.LAMP reaction results were detected using turbidity meter or visual method.The specificity of the constructed method was evaluated by detecting Y.pestis and its closely-related bacteria.The different dilution DNA template was detected with LAMP and PCR to evaluate the sensitivity of the method.Results Thirty strains of bacteria closely related to Y.pestis were detected by the constructed LAMP,and all the results were negative,indicating that the method had a very high specificity.The detection sensitivity of this LAMP assay was 20 pg of DNA per reaction,which was ten-fold that of the regular PCR.The detection reaction was completed in 25 min.Conclusion This LAMP method is quick,sensitive, specific and simple,which is expecked to become an effective method for rapid detection of Y.pestis on the scene.

Objective To evaluate the effect of self-management education on gastroesophageal reflux asthma .Methods 71 gastroesophageal reflux asthma patients were divided into two groups , the management group (n=35) and the control group (n=36).The control group received routine drug therapy and routine outpatient guidance, while the management group was given routine drug treatment and systematic outpatient self -management education.The self-management lasted for 6 months.The symptoms of gastroesophageal reflux , control level of asthma and the pulmonary ventilation function were compared between the two groups .Results After the intervention, the score of symptoms of gastroesophageal reflux in the management group was (5.1 ±1.2), which was significantly lower than (6.8 ±1.6) in the control group (t=5.07,P<0.01).The score of asthma control in the management group was (23.5 ±0.6), which was significantly lower than (22.7 ±0.9) in the control group (t=4.41,P<0.01).On the aspects of pulmonary ventilation function , the 1 second forced vital capacity of the expected percentage , the 1 second forced expiratory rate and peek expiratory flow rate were (87.4 ± 3.9)%,(86.7 ±3.1)%and (8.8 ±1.8) L/s in the management group , which were significantly higher than (83.4 ±4.6)%,(84.3 ±4.2)% and (7.7 ±1.2) L/s in the control group (t =3.96,2.74,3.05, respectively;P<0.01).Conclusions The systematic outpatient self-management education can improve the ability of self care , decrease patients'symptoms and improve patients pulmonary ventilation function .