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Objective:To compare the diagnostic efficacy of Chinese diagnostic model, the 1990 American College of Rheumatology (ACR) classification criteria and the 2018 ACR new classification criteria (draft) for Takayasu arteritis (TA).Methods:A total of 196 TA patients who came to our hospital from January 1, 2009 to May 31, 2019 in the TA database of the department of rheumatology and immunology, Zhongshan Hospital, Fudan University and 131 patients with other vascular diseases visited during the same period were selected. General characteristics, clinical data, laboratory tests and imaging tests of all patients were collected. Categorical variables were presented as numbers and percentages, between-group differences were analyzed using the χ2 test. Continuous variables were presented as the Mean± SD for a normal distribution or median and interquartile range (IQR) for a non-normal distribution. Between-group differences were analyzed using the Student's t-test or Mann- Whitney test, as appropriate. The sensitivity, specificity, positive predictive value, negative predictive value, accuracy and receiver operating characteristics of the above diagnostic/classification criteria area under the curve were analyzed. P<0.05 was considered significant. Results:In terms of sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and area under receiver operating characteristic curve (ROC), Chinese diagnostic models was 85.7%, 96.2%, 89.9%, 97.1%, 81.5%, 0.909, 1990 ACR criteria was 47.4%, 97.7%, 67.6%, 96.9%, 55.4%, 0.726, 2018 ACR classification criteria was 79.1%, 98.5%, 86.5%, 98.7%, 75.9%, 0.888. The difference between the Chinese diagnostic model and the 2018 ACR criteria in AUC was not statistically significant ( Z=1.186 , P>0.05). The sensitivity, accuracy and diagnostic efficiency of Chinese diagnostic model was the best, that of the 1990 ACR classification criteria was the worst, and the specificity of the 2018 ACR classification criteria was the highest. The Kappa value of the 2018 ACR classification criteria and the Chinese diagnostic model was 0.719, which had good consistency, and the Kappa value of the consistency between the 1990 ACR classification criteria and the Chinese TA diagnostic model was 0.516. Conclusion:The Chinese diagnostic model, which is based on the clinical characteristics of the Chinese TA population, has a good diagnostic efficacy for the Chinese population. The 2018 ACR classification criteria (draft) is highly consistent with the Chinese TA diagnostic model, and can be promoted and applied in practice.
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Objective:To analyze the characteristics of patients with Takayasu arteritis (TA) in the east China Takayasu arteritis (ECTA) cohort and their subgroups, and evaluate the disease characteristics.Methods:Patients diagnosed with TA in ECTA cohort from January 2009 to October 2019 were enrolled and their data were analyzed. The characteristics were analyzed and compared within subgroups using t-test or Wilcoxon rank sum test or Chi-square test. Results:A total of 454 patients were included, with the male to female ratio of 1∶4.75(79/375), and the main complaint were dizziness/headache, fatigue, and chest tightness/pain. The type Ⅴ and Ⅰ were the most common angiographic pattern, among which the subclavian artery and carotid artery were most vulnerable, manifested as vascular stenosis. Hypertension, tuberculosis and hepatitis B were common complications. In subgroup comparison, symptoms and inflammation index were much more evident in the active group, female group, <40 years old, and newly diagnosed group. C-reactive protein (CRP)[10(2, 33) mg/L vs 3(1, 14) mg/L, Z=-4.49, P<0.01), erythrocyte sedimentation rate (ESR) [(45±33) mm/1 h vs (25±23) mm/1 h, t=-5.82, P<0.01), in the active group were significantly higher than those in the inactive group, while the ESR in female patients was only higher than that in males, but without statistical significant difference. SAA in the young age group, ESR in the newly diagnosed group was significantly higher than that in the other subgroups [19(6, 95) mg/L vs 10(4, 39) mg/L, Z=2.06, P<0.05] [(44±34) mm/1 h vs (32±28) mm/1 h, t=3.77, P<0.01]. Conclusion:The TA patients are mainly young women, and are in active disease when first being diagnosed. The type Ⅴ and Ⅰ are the most common artery involve-ment pattern. Hypertension and tuberculosis are the most frequent complications.
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Objective To explore the risk factors of pulmonary artery hypertension (PAH) and the its relationship with T cell subsets to provide a foundation for the prevention and treatment of PAH.Methods 154 maintained hemodialysis (MHD) patients in our dialysis center were recruited according to the criterion and divided into two groups subsequently:PAH group (pulmonary artery systolic pressure,PASP ＞ 35 mmHg) and non-PAH group (PASP≤35 mmHg).The related clinical,biochemical and ultrasonic cardiogram data were collected and peripheral blood was acquired to detect the expressions of the surface antigen CD3,CD4,CD8 and CD69 with flow cytometry.Logistic regression analysis was used to find out the relationship between PAH and T cell subsets.Results There was no significant difference between 56 cases of PAH and 98 cases of non-PAH as regards gender,age,mean systolic and diastolic pressure,dialysis durations,morbidities of hypertension and diabetes,smoking rate,and left ventricular diameter.Compared with the non-PAH group,the PAH group demonstrated a lower percent of CD8 T cells and CD8 CD69 T cells,but a much higher left atrial diameter (LAD),Interventricular septum thickness,left ventricular posterior wall thickness,and NT-proBNP.The percentage of T cells,CD4 T cells and CD4 CD69 T cells showed no difference between the two groups.Multivariate analysis confirmed that PAH was negatively independently associated with the percentage of CD8 T cells and CD8CD69 T cells.Conclusions The decreased percentage of CD8 T cells and CD8CD69 T cells in the peripheral blood is a risk factor of PAH in maintained hemodialysis patients,and CD8 T cells may play an important role in the genesis of PAH.
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Objective To investigate the role of T helper 17 cells/interleukin?17(Th17/IL?17) axis in the occurrence of vaginal candidiasis in mice. Methods A total of 120 female BALB/c mice were randomly and equally divided into Ei, En, Ci and Cn groups. Three days before vaginal inoculation, estrogen (Ei and En)groups and control(Ci and Cn)groups received subcutaneous injection of 0.05 mg estradiol and 0.1 ml sterilized soybean oil at the hind legs, respectively, and then the hormone treatment continued every other day until the end of experiment. Infected(Ei and Ci)groups and noninfected(En and Cn) groups were inoculated intravaginally with 10μl(5 × 104 conidia)of Candida albicans suspension and 10μl of sterilized phosphate?buffered saline, respectively. Ten mice were randomly selected from each group and sacrificed on day 3, 7 and 14 after inoculation. The intact vagina tissues were resected and then frozen in liquid nitrogen or embedded in paraffin. Real?time fluorescence?based quantitative PCR(qRT?PCR)and immunofluorescent staining were performed to measure mRNA expression and immunofluorescence intensities of retinoic acid?related orphan receptorγt(RORγt), RORα and IL?17, respectively. Western blot analysis was conducted to determine protein expression of RORγt and IL?17. Results Laser scanning confocal microscopy showed that RORγt, RORα and IL?17 immunofluorescence was mainly located at inflammatory cells of the lamina propria and blood vessels in En and Cn groups, at mucosal epithelium, adherent hyphae, and inflammatory cells of the lamina propria and blood vessels in Ci group, and at mucosal epithelium, vaginal canal and endocytosed hyphae, and inflammatory cells of the lamina propria and blood vessels in Ei group. qRT?PCR and immunofluorescent staining uncovered that mRNA expression and immunofluorescence intensities of RORγt, RORα and IL?17 were significantly higher in En, Ci and Ei groups than in Cn group at the same time points(all P0.05). Moreover, RORγt and IL?17 protein expression in Ci and Ei groups was obviously up?regulated on day 7 after inoculation (RORγt: F = 13.137, P < 0.001; IL?17: F = 11.182, P < 0.001), but was not increased further on day 14. Conclusion Vaginal candida infection can up?regulate the expression of RORγt, RORα and IL?17, suggesting that Th17/IL?17 axis may be involved in the occurrence of vaginal candidiasis in BALB/c mice.
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Objective To evaluate the effects of progesterone on polymorphonuclear leukocyte (PMN)-mediated inflammatory responses to gonococcal infection. Methods Peripheral neutrophils were isolated from heparinized peripheral blood obtained from normal individuals, then divided into 4 groups: progesterone group (pretreated with progesterone only), gonococcus group (stimulated with gonococcal suspension), intervention group (pretreated with progesterone followed by stimuation with gonococcal suspension), and control group (receiving no pretreatment or stimulation). Real-time RT-PCR was conducted to detect the mRNA expression levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)in neutrophils from all groups at 0, 3, 8, 12 and 24 hours after the last treatment, and iNOS protein levels were measured by Western-blot in gonococcus group and intervention group. Results Real-time RT-PCR indicated that the expression levels of iNOS, TNF-α and IL-1β mRNA increased in gonococcus group and intervention group, and reached their peak at 8 hours in gonococcus group, while no significant changes were noted in the above parameters in progesterone group or control group. Also, the level of iNOS, TNF-α and IL-1β mRNA was lower in intervention group than that in the gonococcus group (P ＜ 0.05). Western blot showed an elevation in iNOS protein expression in both gonococcus group and intervention group, and the former group was higher than the latter group in the parameter (P ＜ 0.05). Conclusions Progesterone can downregulate the expressions of iNOS, TNF-α and IL-1 β by PMNs, inhibit the PMN-induced inflammatory responses induced by gonococcal infection, which is likely to be associated with the asymptomatic gonococcal infection in women.
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Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. However, emergence of drug resistance limits its potential use. Plumbagin is a natural quinonoid compound isolated from plant. In this study, induced apoptosis effect of the combined treatment with plumbagin and TRAIL on human melanoma A375 cell line was examined and possible mechanism was investigated. The cells were divided into four groups: control group, plumbagin group (plumbagin, 5 or 10 mumol/L), TRAIL group (TRAIL, 30 ng/mL) and plumbagin+TRAIL group (combined treatment group). The apoptosis, and the expression of DR4 and DR5 were detected by flow cytometry. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that the apoptosis rate was 8.3% in TRAIL group, 10.35%-16.94% in plumbagin group and 52.39%-65.39% in combined treatment group, respectively, with the difference being significant between combined treatment group and plumbagin or TRAIL group (P<0.05 for each). Moreover, plumbagin alone could markedly up-regulate DR5 mRNA and protein expression, and slightly increase DR4 mRNA and protein expression. Treatment of human melanoma A375 cells with plumbagin resulted in the activation of Caspase-3, but not Caspase-8. These results suggest that plumbagin might be useful for TRAIL-based treatment for melanoma.
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Progesterone has nongenomic effects on inducible nitric oxide synthase (iNOS), which is mediated by mitogen activated protein kinase (MAPK) pathways. This effect is supposed to have some potential association with asymptomatic gonococcal infections in women by immunological depression. In this study, polymorphonuclear leukocytes (PMNs) challenged by gonococci were used to study the nongenomic effects of progesterone. The activation of iNOS was assessed by measuring [(3)H] L-arginine converses to [(3)H] L-citrulline, and the activity of MAPK was detected by Western blot. It was found that the activity of iNOS and the yields of NO were enhanced significantly in gonococci-challenged PMNs compared with the controls (P0.05). It was also found subsequently that in the serum specimens collected from gonococci-infected but asymptomatic women, the progesterone level was higher than that in women with severe symptoms (P<0.01). Moreover, the yield of NO had an inverse correlation with progesterone. With these results it suggested that the rapid nongenomic effects of progesterone may inhibit iNOS activation and NO yields mediated by P38MAPK pathways, which were supposed to be concerned with asymptomatic women infected with gonococci.
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Objective: To construct the insulin-like growth factor binding protein 7 (IGFBP7) expression plasmid (pEGFC1-IGFBP7) and to investigate the effect of IGFBP7 on the apoptosis of SK-MEL-28 (human malignant melanoma cell line) cells. Methods: The pEGFC1-IGFBP7 plasmid was constructed; pEGFC1-IGFBP7 and empty plasmids were transfected into SK-MEL-28 cells separately. The transfection efficiency was observed under fluorescence microscope. Apoptosis of SK-MEL-28 cells after transfection was detected by Annexin-FITC/PI staining. Results: The pEGFC1-IGFBP7 plasmid was successfully constructed and was effectively transfected into SK-MEL-28 cells by Effectene reagent, with the transfection rate being 61%. The results of flow cytometry showed that pEGFC1-IGFBP7 significantly induced apoptosis of SK-MEL-28 cells, with the apoptotic rates of pEGFC1-IGFBP7, empty vector, and non-transfected plasmid groups being (28.4±2.57)%, (5.8±0.44)%, and (6.4±0.71)% 24 h after transfection, respectively (F=406.138, P<0.05). Conclusion: pEGFC1-IGFBP7 can effectively induce apoptosis of malignant melanoma SK-MEL-28 cells, which provides an experimental basis for IGFBP7 gene-based therapy of malignant melanoma.
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Objective To study the relationship of symptoms of female gonococcal infections to Chlamydia trachomatis infection, serum sex hormone levels, etc. Methods A total of 136 gonorrhea female patients without obvious symptoms were recruited in this study together with 45 gonorrhea patients with obvious symptoms as the controls. Serum progesterone (P) and estradiol (E2) levels were measured by radio immunoassay (RIA). Cervical swabs were obtained from the subjects and eluted into isotonic saline solution, the elution was divided into 2 portions and tested for the levels of TNF-α and IL-1β by ELISA and for the DNA of C. Trachomatis and N. Gonorrhea with PCR. Statistical analysis was carried out by SPSS for Windows (version 12.0). Results There was no statistical correlation between C. Trachomatis infection and asymptomatic status of female gonococcal infection (χ2 = 0.016, P > 0.05). However, the decrease in the level of TNF-α and IL-1β significantly correlated with the increase in serum progestogen (r = -0.8798, -0.8935, respectively, both P < 0.01). Conclusion The high serum level of progesterone may be associated with the asymptomatic status of gonococcal infection.
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To identify the genomic species of Neisseria gonorrhoeae, evaluate the difference between two molecular epidemiological methods and examine the relationship between sex partners and genotypes of bacteria, 24 strains of Neisseria gonorrhoeae isolated from the outpatients with gonorrhea were identified by using the Opa genotyping and NG-MAST genotyping and the relationship between genotypes and phenotypes was studied. Twenty-four strains of Neisseria gonorrhoeae fell into 10 ST genotypes by NG-MAST genotyping, whereas these strains were classified into 12 OT Opa genotypes by Opa genotyping. A new epidemic strain of ST genotype (217-86% homologisation 178) in China was identified. It is concluded that genotypes of each pair of strains from a pair of patient/ sex partner besides 45/46 are the same, indicating that contagious infection take place between patient and the sex partner. Opa genotyping was more effective than NG-MAST genotyping in identifying the genomic species of Neisseria gonorrhoeae. ST genotype could be further classified into different Opa-types.
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Objective To compare the potentiality of opa typing versus Neisseria gonorrhoeae multi-antigen sequence typing (Ng-MAST) in discrimination of N. gonorrhoeae isolates, and to investigate the consistency of genotypes of N. gonorrhoeae between patients and their sex partners. Methods N. gonor-rhoeae was isolated from patients and their sex partners who attended sexually transmitted disease clinics of the Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from February 2006 to August 2007. All isolates were cultured on Thayer-Martin agar and GC-based choco-late agar, and identified by Gram stain and oxidase reaction. Totally, twenty-four strains of N. gonorrhoeae were isolated form 12 pairs of male-female sex partners. Genomic DNA was isolated from the strains, and subjected to PCR amplification of opa gene as well as pot and tbpB genes, opa typing and Ng-MAST typing were utilized to differentiate the 24 strains respectively. Results The 24 isolates were classified into 10 sequence types (ST) by Ng-MAST typing, and 12 opa types (OT) by opa typing. A novel genotype identi-fied by Ng-MAST (217-86% homologisation 178), was unique in China. Conclusions Both ST and OT of N. gonorrhoeae strains are identical between patients and their corresponding partners except for one pair of partners, 45/46, suggesting that gonorrhea is transmitted to each other by contiguity between sex partners. Opa typing is likely to be more potential than Ng-MAST in genotyping of N.gonorrhoeae strains.
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Objective To study the relationship between cell apoptosis and mRNA expression of c-fos and BNIPl in cutaneous squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). Methods Cell apoptosis and mRNA expression of c-fos and BNIP1 in 48 SCCs and 41 BCCs were determined by TUNEL and in situ hybridization, respectively. Results Apoptosis index (AI) and c-fos mRNA expression in SCCs were higher than those in BCCs (P 0.05). AI was significantly higher in well-differentiated SCCs than in poorly differentiated SCCs (P