Objective:To investigate variation and correlation of type 2 intrinsic lymphocyte(ILC2)and related factors in acute and chronic brucellosis,to identify immune role of ILC2 in chronic brucellosis.Methods:Forty-three patients with acute brucel-losis and 45 patients with chronic brucellosis were compared with 49 healthy controls.ILC2 level in each group was detected by flow cytometry.mRNA level of GATA3 in PBMC was detected by fluorescence quantitative PCR.Serum IL-33,ST2,IL-4 and IL-13 levels were detected by ELISA.ALT and AST levels were measured by fully automatic biochemical analyzer.Results:ILC2 level,GATA3 mRNA in PBMC,serum IL-33,IL-4 and IL-13 levels in chronic brucellosis group were significantly higher than healthy control group and acute brucellosis group(P<0.01).Correlation analysis showed that ILC2 level was positively correlated with GATA3 mRNA,IL-33,IL-4 and IL-13 levels,while negatively correlated with ST2 level.ROC curve suggested that ILC2,GATA3,IL-33 and ST2 had good predictive ability for severity of brucellosis patients(AUC>0.7).Conclusion:Increase of ILC2 and its related factors are closely related to chronic brucella infection,which may play an important immune role in pathogenesis of brucella infection.

To investigate the pharmacological substances basis and anti-vitiligo mechanism of Carum carvi L. (caraway) fruits, chemical and blood-absorbed components were identified using mass spectrometry combined with literature study and database analysis. A “blood-absorbed components–target genes–pathways” network was constructed through network pharmacology. The pharmacological effects of Carum carvi L. extract and its key blood-absorbed component, acacetin, were validated in vitro. 72 chemical components were identified in the extract, with 11 prototype blood-absorbed components and 26 metabolites being detected in the plasma of SD rats. 14 key active components and 24 key targets were predicted. In vitro experiments demonstrated that acacetin at 10 μmol/L exhibited melanogenesis-promoting and tyrosinase-activating effects compared with the positive control, significantly upregulating the expression of microphthalmia-associated transcription factor (MITF) and tyrosinase (tyrosinase, TYR). This study comprehensively analyzes the chemical and blood-absorbed components of Carum carvi L. and elucidates its pharmacological substances basis, which provides a theoretical foundation for the anti-vitiligo effects of acacetin.

Objective:To establish a high performance liquid chromatography-evaporative light scattering detector(HPLC-ELSD)technique for simultaneous determining the content of excipients sucrose and mannitol in meningo-coccal polysaccharide vaccine.Methods:Using NanoChrom Sugar-10Ca analytical column(300 mm × 7.8 mm)and HPLC system(Agilent 1260).With purified water as the mobile phase,a flow rate of 0.5 mL·min-1,column temperature was 80 ℃,and the injection volume was 50 μL.The evaporative light detector was based on nitrogen.The carrier gas flow rate is 3.2 L·min-1,the temperature of drift tube was 1 10 ℃,the gain value was 1,and the impactor was"mode 1".This assay was subsequently validated for its system suitability,specificity,repeatability,intermediate precision and linearity,and accuracy.The established method was used to assay the contents of the sucrose and mannitol in four batches of meningococcal polysaccharide vaccine.Results:The estab-lished HPLC-ELSD method showed good systemic suitability.Specificity validation showed that there was no inter-ference peak in the blank solvent;the separation of the target peaks between sucrose and mannitol was＞2.0.The relative standard deviations(RSD)value of peak area of sucrose and mannitol in six tests were 0.44％and 0.38％,respectively.RSD of intermediate precision of both sucrose and mannitol were lower than 2.00％,indica-ting that the precision of high performance liquid chromatography instrument was well.The linear range of two excipients were 12.5-150.0 μg·mL-1(R2＞0.99,respectively).The recovery rate of sucrose and mannitol were 95.74％-99.33％,94.37％-98.85％,respectively.There was no significant difference in the contents of sucrose and mannitol in 4 batches of meningococcal polysaccharide vaccine.Conclusion:The HPLC-ELSD method showed good specificity,precision,linearity and accuracy,and the test results were stable and reliable,so that it is suitable for simultaneous determination of sucrose and mannitol contents of meningococcal polysaccharide injections.

Objective:To investigate variation and correlation of type 2 intrinsic lymphocyte(ILC2)and related factors in acute and chronic brucellosis,to identify immune role of ILC2 in chronic brucellosis.Methods:Forty-three patients with acute brucel-losis and 45 patients with chronic brucellosis were compared with 49 healthy controls.ILC2 level in each group was detected by flow cytometry.mRNA level of GATA3 in PBMC was detected by fluorescence quantitative PCR.Serum IL-33,ST2,IL-4 and IL-13 levels were detected by ELISA.ALT and AST levels were measured by fully automatic biochemical analyzer.Results:ILC2 level,GATA3 mRNA in PBMC,serum IL-33,IL-4 and IL-13 levels in chronic brucellosis group were significantly higher than healthy control group and acute brucellosis group(P<0.01).Correlation analysis showed that ILC2 level was positively correlated with GATA3 mRNA,IL-33,IL-4 and IL-13 levels,while negatively correlated with ST2 level.ROC curve suggested that ILC2,GATA3,IL-33 and ST2 had good predictive ability for severity of brucellosis patients(AUC>0.7).Conclusion:Increase of ILC2 and its related factors are closely related to chronic brucella infection,which may play an important immune role in pathogenesis of brucella infection.

Objective:To establish a high performance liquid chromatography-evaporative light scattering detector(HPLC-ELSD)technique for simultaneous determining the content of excipients sucrose and mannitol in meningo-coccal polysaccharide vaccine.Methods:Using NanoChrom Sugar-10Ca analytical column(300 mm × 7.8 mm)and HPLC system(Agilent 1260).With purified water as the mobile phase,a flow rate of 0.5 mL·min-1,column temperature was 80 ℃,and the injection volume was 50 μL.The evaporative light detector was based on nitrogen.The carrier gas flow rate is 3.2 L·min-1,the temperature of drift tube was 1 10 ℃,the gain value was 1,and the impactor was"mode 1".This assay was subsequently validated for its system suitability,specificity,repeatability,intermediate precision and linearity,and accuracy.The established method was used to assay the contents of the sucrose and mannitol in four batches of meningococcal polysaccharide vaccine.Results:The estab-lished HPLC-ELSD method showed good systemic suitability.Specificity validation showed that there was no inter-ference peak in the blank solvent;the separation of the target peaks between sucrose and mannitol was＞2.0.The relative standard deviations(RSD)value of peak area of sucrose and mannitol in six tests were 0.44％and 0.38％,respectively.RSD of intermediate precision of both sucrose and mannitol were lower than 2.00％,indica-ting that the precision of high performance liquid chromatography instrument was well.The linear range of two excipients were 12.5-150.0 μg·mL-1(R2＞0.99,respectively).The recovery rate of sucrose and mannitol were 95.74％-99.33％,94.37％-98.85％,respectively.There was no significant difference in the contents of sucrose and mannitol in 4 batches of meningococcal polysaccharide vaccine.Conclusion:The HPLC-ELSD method showed good specificity,precision,linearity and accuracy,and the test results were stable and reliable,so that it is suitable for simultaneous determination of sucrose and mannitol contents of meningococcal polysaccharide injections.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.

Objective:To explore expression of Siglec-7 on monocytes in peripheral blood of patients with brucellosis and cor-relation between Siglec-7+monocytes and Th1/Th2 cells,and to analyze clinical significance of Siglec-7 molecule in patients with bru-cellosis.Methods:Fifty patients with newly diagnosed Brucella infection(BI group)and 46 healthy controls(control group)were in-cluded.Flow cytometry was used to detect expression of Siglec-7 on monocytes,and correlation between Siglec-7+monocytes and clini-cal indicators of patients with brucellosis were analyzed.Flow cytometry was used to detect Th1/Th2 cells levels,and CBA method was used to detect IFN-γ and IL-4 in peripheral serum.Correlation between Siglec-7+monocytes and Th1/Th2 cells,IFN-γ/IL-4 were ana-lyzed.Results:Siglec-7+monocytes level in BI group was significantly higher than that in control group(P<0.001);Siglec-7+mono-cytes level in patients with abnormal liver function and lung X-ray were higher than those in patients with normal liver function and lung X-ray(P<0.001,P<0.05);Compared with control group,Th2 cell and IL-4 levels were significantly increased(P<0.05),while Th1 cells and IFN-γ levels were significantly reduced(P<0.001);Th1/Th2 and IFN-γ/IL-4 were also significantly reduced(P<0.001).Siglec-7+monocyte level was negatively correlated with Th1 and IFN-γ(r=-0.651,r=-0.407),while was positively correlated with Th2 and IL-4(r=0.706,r=0.530).Conclusion:During Brucella infection,increased percentage of Siglec-7+monocytes may be involved in Th1/Th2 cell imbalance.

Objective:pH low insertion peptide (pHLIP)-variant 7 (var7)-fluorescein isothiocyanate (FITC) was used to explore an accurate imaging tool that targeted burn wounds to better perform burn debridement.Methods:Twelve rat models of burn wound were established and pHLIP-var7-FITC with different concentrations (0.5, 1.5 and 2.0 mg/ml) were injected from the rat tail vein for in vivo fluorescence imaging. By determining the concentration of fluorescent conjugates to the burn wound, the scope of wound injury necrosis was judged by combining pathological sections, and its residue and toxicity in important organs such as heart, liver, kidneys, and brain were detected. The Kruskal-Wallis rank sum test, Bonferroni correction method and one-way analysis of variance were used for data analysis. Results:Within 24 h, the fluorescence photons per unit area of the burn wound in the group of 0.5 mg/ml, 1.5 mg/ml and 2.0 mg/ml were 1.49(1.31, 1.65), 2.46(1.88, 2.68), 2.77 (1.94, 3.10)×10 7 p·s -1·cm -2·Sr -1, with significant differences in the overall distribution of fluorescence photons ( H=73.55, P<0.001). The fluorescence intensity was stronger in the group with higher concentration, but with no significant difference in the number of fluorescence photons between the group of 1.5 mg/ml and 2.0 mg/ml ( P=0.263, Bonferroni correction method). At 14 time points (0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, 12, 24 h), there was no significant difference in the overall mean of fluorescence photons ( F=1.04, P=0.419), and the tissue with burn necrosis seen in tissue sections was highly consistent with the fluorescence imaging region. There was no obvious fluorescence residue in the heart, liver, kidney and brain sections. Conclusion:In superficial second-degree burn tissue, pHLIP-var7-FITC can accurately target and gather on the burn wound within 24 h, showing a clear boundary between burn tissue and normal tissue, which can assist clinical surgical debridement to determine the extent of injury.

Objective To investigate the interventional effect and mechanism of a novel ampelopsis hydrogel on dampness-heat accumulation syndrome of gouty arthritis. Methods A total of 90 patients with gouty arthritis who met the diagnostic criteria of western medicine and were differentiated as damp-heat accumulation syndrome of traditional Chinese medicine(TCM) were randomly divided into treatment group, control group and blank group, with 30 patients in each group. The blank group was treated with etoricoxib only, the control group was treated with etoricoxib combined with ampelopsis hydrogel, and the treatment group was treated with etoricoxib combined with external application of ampelopsis hydrogel. The clinical efficacy, time to symptom improvement, safety, comfort, changes in syndrome scores of TCM, serum inflammatory factors[C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), erythrocyte sedimentation rate (ESR)], NF-κB signaling pathway-related proteins, Visual Analogue Scale (VAS) scores for pain, and joint mobility were compared among the three groups before and after treatment. Results The total effective rates in the treatment group and control group were 93.33% and 90.00%, respectively, which were higher than 70.00% in the blank group (P < 0.05). The time for improvement of pain, redness, tenderness, and limited joint mobility in the treatment group was shorter than that in the control group and blank group (P < 0.05). After 7 days of treatment, the TCM syndrome score in the treatment group was lower than that in the control group and blank group, and the levels of serum CRP, TNF-α, and ESR and the expressions of NF-κB signalingpathway-related proteins P50 and P65 in the treatment group and control group were lower than those in the blank group (P < 0.05). After 7 days of treatment, the VAS score in the treatment group was lower than that in the control group and blank group, and the comfort score in the treatment group was higher than that in the control group (P < 0.05). There was no significant difference in the incidence of adverse reactions among the three groups (P>0.05). Conclusion The effect of ampelopsis hydrogel in treating gouty arthritis is better than that of ampelopsis paste, and its mechanism may be related to the regulation of the NF-κB pathway and inhibition of inflammatory factor expression. The hydrogel is easy to use, hygienic, and comfortable, and is expected to become a safe, effective, and convenient external medicine for gouty arthritis.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.