Objective:To learn about the levels of 25-hydroxy vitamin D (25-OH VD), TBNK lymphocyte subsets, and cytokines in peripheral blood of patients with brucellosis.Methods:A prospective design was adopted, one hundred patients with brucellosis admitted to the Department of Infectious Diseases, Beidahuang Industry Group General Hospital from May 2024 to February 2025 were selected as the brucellosis group, and one hundred healthy individuals who underwent physical examinations at the hospital during the same period were selected as the control group. The peripheral blood 25-OH VD levels were detected by chemiluminescence method. Further, 100 patients with brucellosis were divided into a brucellosis combined with osteoarthritis group (74 cases) and a brucellosis without osteoarthritis group (26 cases). Flow cytometry was used to detect the counts of peripheral blood TBNK lymphocyte subsets and cytokine levels. Meanwhile, Spearman rank correlation was used to analyze the correlation between peripheral blood 25-OH VD levels and TBNK lymphocyte subsets counts as well as cytokine levels in patients with brucellosis complicated by osteoarthritis.Results:The peripheral blood 25-OH VD level in the brucellosis group [20.31 (15.74, 24.35) ng/ml] was significantly lower than that of the control group [25.18 (21.13, 29.59) ng/ml], and the difference was statistically significant ( Z = - 5.07, P < 0.001). The peripheral blood 25-OH VD level [18.05 (13.79, 23.74) vs 22.43 (19.93, 28.25) ng/ml], CD4 + T cell count [(860 ± 275) vs (1 036 ± 376) cells/μl], and interleukin (IL)-6 levels [4.17 (2.14, 9.41) vs 7.83 (5.97, 11.34) ng/L] in the brucellosis combined with osteoarthritis group were significantly lower than those in the brucellosis without osteoarthritis group ( Z/t = - 2.88, 2.20, - 2.85, P = 0.004, 0.035, 0.004). Correlation analysis showed that the peripheral blood 25-OH VD level in patients with brucellosis complicated by osteoarthritis was positively correlated with the counts of CD45 +, CD3 + T, CD4 + T, CD8 + T, and natural killer cells ( r = 0.31, 0.26, 0.25, 0.25, 0.25, P = 0.007, 0.027, 0.032, 0.031, 0.032), and negatively correlated with IL-17A level ( r = - 0.40, P < 0.001). Conclusion:Patients with brucellosis have insufficient 25-OH VD, and those with osteoarthritis have lower 25-OH VD level, CD4 + T cell count, and IL-6 level than those without osteoarthritis.

Objective:To investigate the application in evaluating the course and the clinical effects of serum cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) levels, as well as neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) in patients with brucellosis.Methods:Using case-control method, from February 2023 to March 2024, 274 confirmed brucellosis patients [divided into acute and chronic groups ( n = 165, 109) according to the course of the disease] and 70 healthy individuals (control group) were selected at Beidahuang Group General Hospital for serum cytokines detection using cytometric bead array (CBA) method. Blood routine test, serum agglutination test (SAT) and blood culture were performed at the same time, and NLR and PLR were calculated. Cytokine levels, NLR, and PLR were compared in patients with different disease duration, with or without complications, with different SAT titers [high ( > 1 ∶ 100) and low (≤1 ∶ 100)], and with different blood culture results, and the effects of each indicator on the course of brucellosis were analyzed by logistic regression. Results:The levels of IL-2, IL-6, IL-10, IL-17, TNF-α and IFN-γ in the acute group [ M ( Q1, Q3): 0.32 (0.15, 0.70), 18.97 (10.70, 36.86), 2.54 (1.49, 4.36), 1.41 (0.38, 3.05), 1.31 (0.77, 2.33), 11.60 (2.30, 36.75) ng/L] were higher than those in the chronic group [0.18 (0.06, 0.43), 1.68 (0.75, 5.74), 0.88 (0.40, 1.93), 0.29 (0.09, 0.87), 0.59 (0.31, 1.07), 0.72 (0.33, 1.42) ng/L] and control group [0.10 (0.05, 0.30), 1.52 (0.09, 2.80), 0.72 (0.35, 1.16), 0.08 (0.03, 0.20), 0.55 (0.20, 0.96), 0.68 (0.41, 1.25) ng/L, P < 0.05]. The IFN-γ level in the group with complications of brucellosis was lower than that in the group without complications, while the NLR and PLR were higher than those in the group without complications ( P < 0.05). The levels of IL-6, IL-17, TNF-α, and IFN-γ in the high titer group were higher than those in the low titer group, and the NLR was lower than that in the low titer group ( P < 0.05). The levels of IFN-γ and TNF-α of blood culture positive patients in the acute group were higher than those of blood culture negative patients ( P < 0.05). Univariate analysis showed that all 7 cytokines could affect the course of brucellosis ( P < 0.05). Multivariate analysis showed that IL-6, TNF-α, and IFN-γ were independent influencing factors of the course of brucellosis [ OR (95% CI) = 0.87 (0.83, 0.91), 0.55 (0.32, 0.97), 0.80 (0.72, 0.88), P < 0.05]. Conclusions:The levels of cytokines IL-2, IL-6, IL-10, IL-17, TNF-α, and IFN-γ can reflect the course progression of brucellosis patients, IL-6, IFN-γ and TNF-α can also serve as independent influencing factors for brucellosis progression. NLR and PLR may become inflammatory markers for predicting Brucella infection.

Objective:To investigate influences of tetrandrine(Tet)on proliferation,apoptosis and immune escape of melanoma cells by regulating cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signal pathway.Methods:MV3 cells were divided into NC group,Tet-L group(5 μmol/L),Tet-M group(10 μmol/L),Tet-H group(15 μmol/L),RU.521(cGAS inhibi-tor)group(1 μmol/L),Tet-H+RU.521 group(15 μmol/L+1 μmol/L).Proliferation of MV3 cells was detected by CCK-8 and plate cloning test;apoptosis of MV3 cells was detected by flow cytometry;Western blot was used to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),TGF-β,cGAS,STING proteins in MV3 cells.Cells in above groups were co-cultured with NK cells for 48 h through Transwell chamber,and named NC co-culture group,Tet-L co-culture group,Tet-M co-culture group,Tet-H co-culture group,RU.521 co-culture group,Tet-H+RU.521 co-culture group,respectively.ELISA was used to detect levels of IFN-γ and Granzyme B in supernatant of co-cultured cells;change of NK cell killing activity was detected.Results:Compared with NC group,A450,cloning rate,PCNA and TGF-β protein expressions of MV3 cells in Tet-L group,Tet-M group and Tet-H group were decreased,apoptosis rate,Bax,cGAS,STING protein expressions were increased,which was in a dose dependent man-ner,change trend of corresponding indexes in RU.521 group was contrary to the above(P<0.05);compared with NC co-culture group,levels of IFN-γ,Granzyme B and NK cell killing activity in supernatant of Tet-L co-culture group,Tet-M co-culture group and Tet-H co-culture group were increased,which was in a dose-dependent manner,change trend of corresponding indexes in RU.521 co-culture group was opposite to the above(P<0.05);RU.521 attenuated inhibition of high-dose Tet on MV3 cell proliferation,immune escape and its promotion on apoptosis.Conclusion:Tet may inhibit melanoma cell proliferation,immune escape and promote cell apoptosis by activating cGAS-STING signal pathway.

Objective:To investigate the application in evaluating the course and the clinical effects of serum cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) levels, as well as neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) in patients with brucellosis.Methods:Using case-control method, from February 2023 to March 2024, 274 confirmed brucellosis patients [divided into acute and chronic groups ( n = 165, 109) according to the course of the disease] and 70 healthy individuals (control group) were selected at Beidahuang Group General Hospital for serum cytokines detection using cytometric bead array (CBA) method. Blood routine test, serum agglutination test (SAT) and blood culture were performed at the same time, and NLR and PLR were calculated. Cytokine levels, NLR, and PLR were compared in patients with different disease duration, with or without complications, with different SAT titers [high ( > 1 ∶ 100) and low (≤1 ∶ 100)], and with different blood culture results, and the effects of each indicator on the course of brucellosis were analyzed by logistic regression. Results:The levels of IL-2, IL-6, IL-10, IL-17, TNF-α and IFN-γ in the acute group [ M ( Q1, Q3): 0.32 (0.15, 0.70), 18.97 (10.70, 36.86), 2.54 (1.49, 4.36), 1.41 (0.38, 3.05), 1.31 (0.77, 2.33), 11.60 (2.30, 36.75) ng/L] were higher than those in the chronic group [0.18 (0.06, 0.43), 1.68 (0.75, 5.74), 0.88 (0.40, 1.93), 0.29 (0.09, 0.87), 0.59 (0.31, 1.07), 0.72 (0.33, 1.42) ng/L] and control group [0.10 (0.05, 0.30), 1.52 (0.09, 2.80), 0.72 (0.35, 1.16), 0.08 (0.03, 0.20), 0.55 (0.20, 0.96), 0.68 (0.41, 1.25) ng/L, P < 0.05]. The IFN-γ level in the group with complications of brucellosis was lower than that in the group without complications, while the NLR and PLR were higher than those in the group without complications ( P < 0.05). The levels of IL-6, IL-17, TNF-α, and IFN-γ in the high titer group were higher than those in the low titer group, and the NLR was lower than that in the low titer group ( P < 0.05). The levels of IFN-γ and TNF-α of blood culture positive patients in the acute group were higher than those of blood culture negative patients ( P < 0.05). Univariate analysis showed that all 7 cytokines could affect the course of brucellosis ( P < 0.05). Multivariate analysis showed that IL-6, TNF-α, and IFN-γ were independent influencing factors of the course of brucellosis [ OR (95% CI) = 0.87 (0.83, 0.91), 0.55 (0.32, 0.97), 0.80 (0.72, 0.88), P < 0.05]. Conclusions:The levels of cytokines IL-2, IL-6, IL-10, IL-17, TNF-α, and IFN-γ can reflect the course progression of brucellosis patients, IL-6, IFN-γ and TNF-α can also serve as independent influencing factors for brucellosis progression. NLR and PLR may become inflammatory markers for predicting Brucella infection.

Objective:To investigate the pulmonary function in patients with Parkinson's disease(PD)and the association between pulmonary function and motor symptoms.Methods:A total of 104 PD patients were included in the study,and a portable spirometer was used to assess pulmonary function.According to the results of pulmonary function,the patients were divided into ventilation dys-function group with 37 patients and non-ventilation dysfunction with 67 patients,and clinical data were compared between the two groups.Univariate and multivariate logistic regression analyses were used to investigate the factors associated with abnormal pulmonary function,and a Spearman correlation analysis was used to investigate the correlation between pulmonary function parameters and motor symptoms.Results:There were significant differences in the scores of bradykinesia and postural gait between the two groups(P<0.05),and the logistic regression analysis showed a positive correlation between pulmonary ventilation dysfunction and postural gait score(regression coefficient β=0.171,95%CI=1.015-1.386,P=0.032).There were significant differences between the patients with different stages of PD in the pulmonary function parameters of the percentage of forced vital capacity in predicted value(t=2.143,P=0.034),the percentage of forced expiratory volume in 1 second(t=2.118,P=0.037),the percentage of predicted peak expiratory flow(t=2.002,P=0.048),maximum expiratory pressure as a percentage of predicted value(t=2.412,P=0.018),and maximum inspiratory pressure as a percentage of predicted value(t=2.119,P=0.036).Conclusion:The pulmonary function parameters of PD patients are lower than expected values,and the incidence rate of pulmonary ventilation dysfunction is 35.58%,with the main manifestation of restrictive ventilatory dysfunction.There is a correlation between pulmonary function and motor symptoms in PD patients.

Objective:To learn about the levels of 25-hydroxy vitamin D (25-OH VD), TBNK lymphocyte subsets, and cytokines in peripheral blood of patients with brucellosis.Methods:A prospective design was adopted, one hundred patients with brucellosis admitted to the Department of Infectious Diseases, Beidahuang Industry Group General Hospital from May 2024 to February 2025 were selected as the brucellosis group, and one hundred healthy individuals who underwent physical examinations at the hospital during the same period were selected as the control group. The peripheral blood 25-OH VD levels were detected by chemiluminescence method. Further, 100 patients with brucellosis were divided into a brucellosis combined with osteoarthritis group (74 cases) and a brucellosis without osteoarthritis group (26 cases). Flow cytometry was used to detect the counts of peripheral blood TBNK lymphocyte subsets and cytokine levels. Meanwhile, Spearman rank correlation was used to analyze the correlation between peripheral blood 25-OH VD levels and TBNK lymphocyte subsets counts as well as cytokine levels in patients with brucellosis complicated by osteoarthritis.Results:The peripheral blood 25-OH VD level in the brucellosis group [20.31 (15.74, 24.35) ng/ml] was significantly lower than that of the control group [25.18 (21.13, 29.59) ng/ml], and the difference was statistically significant ( Z = - 5.07, P < 0.001). The peripheral blood 25-OH VD level [18.05 (13.79, 23.74) vs 22.43 (19.93, 28.25) ng/ml], CD4 + T cell count [(860 ± 275) vs (1 036 ± 376) cells/μl], and interleukin (IL)-6 levels [4.17 (2.14, 9.41) vs 7.83 (5.97, 11.34) ng/L] in the brucellosis combined with osteoarthritis group were significantly lower than those in the brucellosis without osteoarthritis group ( Z/t = - 2.88, 2.20, - 2.85, P = 0.004, 0.035, 0.004). Correlation analysis showed that the peripheral blood 25-OH VD level in patients with brucellosis complicated by osteoarthritis was positively correlated with the counts of CD45 +, CD3 + T, CD4 + T, CD8 + T, and natural killer cells ( r = 0.31, 0.26, 0.25, 0.25, 0.25, P = 0.007, 0.027, 0.032, 0.031, 0.032), and negatively correlated with IL-17A level ( r = - 0.40, P < 0.001). Conclusion:Patients with brucellosis have insufficient 25-OH VD, and those with osteoarthritis have lower 25-OH VD level, CD4 + T cell count, and IL-6 level than those without osteoarthritis.

Objective:To investigate influences of tetrandrine(Tet)on proliferation,apoptosis and immune escape of melanoma cells by regulating cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signal pathway.Methods:MV3 cells were divided into NC group,Tet-L group(5 μmol/L),Tet-M group(10 μmol/L),Tet-H group(15 μmol/L),RU.521(cGAS inhibi-tor)group(1 μmol/L),Tet-H+RU.521 group(15 μmol/L+1 μmol/L).Proliferation of MV3 cells was detected by CCK-8 and plate cloning test;apoptosis of MV3 cells was detected by flow cytometry;Western blot was used to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),TGF-β,cGAS,STING proteins in MV3 cells.Cells in above groups were co-cultured with NK cells for 48 h through Transwell chamber,and named NC co-culture group,Tet-L co-culture group,Tet-M co-culture group,Tet-H co-culture group,RU.521 co-culture group,Tet-H+RU.521 co-culture group,respectively.ELISA was used to detect levels of IFN-γ and Granzyme B in supernatant of co-cultured cells;change of NK cell killing activity was detected.Results:Compared with NC group,A450,cloning rate,PCNA and TGF-β protein expressions of MV3 cells in Tet-L group,Tet-M group and Tet-H group were decreased,apoptosis rate,Bax,cGAS,STING protein expressions were increased,which was in a dose dependent man-ner,change trend of corresponding indexes in RU.521 group was contrary to the above(P<0.05);compared with NC co-culture group,levels of IFN-γ,Granzyme B and NK cell killing activity in supernatant of Tet-L co-culture group,Tet-M co-culture group and Tet-H co-culture group were increased,which was in a dose-dependent manner,change trend of corresponding indexes in RU.521 co-culture group was opposite to the above(P<0.05);RU.521 attenuated inhibition of high-dose Tet on MV3 cell proliferation,immune escape and its promotion on apoptosis.Conclusion:Tet may inhibit melanoma cell proliferation,immune escape and promote cell apoptosis by activating cGAS-STING signal pathway.

Objective: The study aimed to establish humanized patient-derived xenograft (PDX) mouse models of ovarian clear cell carcinoma (OCCC) and evaluate their therapeutic responses. Methods: PDX models and their humanized counterparts (CD34+ humanized PDX models) derived from the same tumor source were developed, and the therapeutic responses were compared between the models. Results: Treatment with a phosphatidylinositol 3-kinase (PI3K) inhibitor significantly reduced tumor size in traditional OCCC PDX models (p=0.021). Although differences in tumor growth between traditional PDX models and humanized PDX models were observed, they were not statistically significant (p=0.438). However, treatment effects of PI3K inhibitor differed significantly between conventional and humanized mice (p=0.006). In the Humanized PDX cohort, both programmed cell death protein-1 antibody monotherapy and PI3K inhibitor treatment slowed tumor growth relative to controls, with a synergistic effect noted during the latter part of the study, though these effects were not statistically significant. Conclusion This pioneering study successfully develop a humanized PDX model for OCCC, highlighting differential responses to treatments compared to conventional PDX models.

Objective:To analyze the clinical efficacy and pregnancy outcomes of gonadotropin-releasing hormone agonist (GnRH-a) based fertility-sparing re-treatment in women with endometrial carcinoma (EC) and atypical endometrial hyperplasia (AEH) who failed with oral progestin therapy.Methods:Forty cases with EC or AEH who failed to respond to oral progestin were included from January 2012 to December 2020 at Peking Union Medical College Hospital. Combination of GnRH-a with levonorgestrel-releasing intrauterine system (group GLI: a subcutaneous injection of GnRH-a every 4 weeks and LNG-IUS insertion constantly) or the combination of GnRH-a with aromatase inhibitor (group GAI: a subcutaneous injection of GnRH-a every 4 weeks and oral letrozole 2.5 mg, daily) were used for these patients. Histological evaluation were performed at the end of each course (every 3-4 months) by hysteroscopy and curettage. After the complete remission (CR), all patients were followed up regularly.Results:(1) Clinical characteristics:among the 40 patients with EC or AEH, the median age at diagnosis was 31 years (range: 22-40 years) and the median body mass index was 24.7 kg/m 2 (range: 18.9-39.5 kg/m 2). (2) Efficacy of fertility-sparing re-treatment: 37 (92%, 37/40) patients achieved CR, 6 (6/7) in AEH and 31 (94%, 31/33) in EC patients. The CR rate was 93% (26/28) and 11/12 in group GLI and GAI, respectively. The median time to CR was 5 months (range: 3-12 months). At the end of the first therapy course, the CR rates in AEH and EC were 5/7 and 42% (14/33), at the second course, the CR rates were 6/7 and 82% (27/33), respectively. (3) Recurrence: after 25 months of median follow-up duration (range: 10-75 months), 8 (22%, 8/37) women developed recurrence, 1/6 in AEH and 7 (23%, 7/31) in EC patients, with the median recurrence time of 18 months (range: 9-26 months). Among them, two cases who had completed childbirth chose to receive hysterectomy directly. Six patients met the criteria of fertility-preserving therapy and received conservative treatment again and 5 (5/6) of them achieved CR. (4) Pregnancy: of the 37 patients with CR, 33 desired to conceive. Ten women attempted to get pregnancy spontaneously and 23 cases with assisted reproductive technology. Fourteen (42%, 14/33) patients became pregnant, including 9 (27%, 9/33) live births, 3 (9%, 3/33) missed abortions, and 2 (6%, 2/33) miscarriages at the second trimester. Conclusions:GnRH-a based fertility-sparing re-treatment in AEH or EC patients who failed with oral progestin therapy achieved good treatment effect and reproductive outcomes. It is an encouraging alternative regime for patients who failed with oral progestin therapy.

Objective:To investigate the clinical significance of six serological methods for diagnosis of brucellosis.Methods:Totally 160 cases of brucellosis patients were selected as the case group and 235 cases of health medical examiners were selected as the control group from January to June 2018 in Heilongjiang Agricultural Reclamation Bureau General Hospital. Six methods were used for detection of brucellosis, including Rose Bengal plate agglutination test (RBPT), fluorescence polarization assay (FPA), indirect enzyme linked immunosorbent assay (iELISA), cysteine agglutination test, standard tube agglutination test (SAT), and anti-globulin test (Coomb's). The consistency of the 4 preliminary screening methods (RBPT vs FPA, iELISA, cysteine agglutination test), and 2 diagnosis methods (SAT vs Coomb's) was compared, and the sensitivity, specificity, rates of missed diagnosis and misdiagnosis were analyzed. Results:In the preliminary screening methods, the consistency of RBPT with FPA and iELISA was relatively consistent (Kappa = 0.872, 0.784), and the consistency with cysteine agglutination test was generally consistent (Kappa = 0.543). In the diagnosis methods, the consistency between SAT and Coomb's was relatively consistent (Kappa = 0.861). The sensitivity of FPA, iELISA, cysteine agglutination test and Coomb's was 91.03%, 75.00%, 56.41% and 80.14%, respectively. The specificity was 95.81%, 100.00%, 98.74% and 100.00%, respectively. The rates of missed diagnosis were 8.97%, 25.00%, 43.59% and 19.86%, respectively. The rates of misdiagnosis were 4.19%, 0, 1.26%, and 0, respectively.Conclusions:FPA and iELISA have high sensitivity and specificity, and are suitable for clinical promotion. The combined detection of multiple serological methods will reduce the probability of misdiagnosis and missed diagnosis of brucellosis, thereby improving the diagnostic level of brucellosis.