1.The expression of macrophage migration inhibitory factor in PBMC,serum and lacrimal fluid of patients with atopic dermatitis and its clinical significance
Lei MA ; Shiyi CHEN ; Yanyang LI ; Chunmei SHU ; Junhua ZHANG
Chinese Journal of Laboratory Medicine 2010;33(12):1155-1159
Objective To investigate the expression of MIF in PBMC, sera and lacrimal fluid samples in patients with AD, and study the diagnostic significance of MIF in AD. Methods Forty-three AD patients (11 mild AD patients,23 moderate AD patients, 9 severe AD patients classfied by SCORAD index)and 31 age- and sex-matched healthy control subjects were recruited. Real-time RT-PCR was employed to analyze the expression of MIF mRNA in PBMC. ELISA was performed to detect the concentrations of MIF in sera and lacrimal fluid samples. Results AD patients had significantly higher levels of MIF mRNA in PBMC than normal controls [7.46 (3.38-8.90) vs 1.67 ( 1.24-2.45 ), Z=-6.141, P < 0.05]. The concentrations of MIF in sera and lacrimal fluid samples in AD patients were also markedly higher than those of normal controls [serum 36. 32( 11.89-43.80) μg/L vs 7.89(6.13-9.54) μg/L, Z = -6.180,P <0.05; lacrimal fluid 12.66(2.01-20.12) μg/L vs 0.85(0.77-1.06) μg/L, Z = -4.118,P <0.05]. MIF mRNA levels were 2.35 ( 2.12-2.49 ) , 7.83 ( 6. 54-8.90 ) and 8.76 ( 8.22-9.73 ) in mild, moderate and severe AD respectively, and the expression was higher in moderate and severe AD than in normal controls (Z= -6.237, -4.520,P <0.05). MIF serum concentrations were 8.98(7.90-10.51) μg/L, 36.50 (29.78-43.23) μg/L and 45.70(41.27-48. 84) μg/L in mild, moderate and severe AD respectively, and the differences were significant in moderate and severe AD compared to normal controls ( Z = - 6.238,- 4.521, P < 0.05 ). The lacrimal fluid MIF oncentrations were 1.10 ( 0.83-1.35 ) μg/L, 12.66 ( 9.76-15.87) μg/L and 24. 65 ( 19.29-30.94) μg/L in mild, moderate and severe AD respectively. Similarly,they increased significantly in moderate and severe AD compared to normal controls (Z = -4.062,- 3.372, P < 0.05 ). In moderate and severe AD, MIF mRNA levels in PBMCs and MIF concentrations in sera and lacrimal fluid samples were all positively correlated with the severity of AD ( r = 0.395, 0.404,0.515, P < 0.05 ). Conclusions The expression of MIF in PBMCs, sera and lacrimal fluid samples is higher in different course of AD. MIF can serve as a useful laboratory parameter for evaluation of AD activity and severity.
2.Expression of miR-155 in peripheral blood and skin lesions from as well as its relationship with Th17 cells in patients with atopic dermatitis
Lei MA ; Haibo XUE ; Xiuhao GUAN ; Chunmei SHU ; Juan YU ; Junhua ZHANG ; Beilei YUN
Chinese Journal of Dermatology 2014;47(1):15-18
Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78% ± 0.52% vs.0.47% ± 0.15%,P< 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P< 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P< 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P< 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P< 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P < 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P < 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P < 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.
3.Value of mesenteric CT angiography in diagnosis of small intestinal neoplasms
Meihong SHENG ; Shenchu GONG ; Bosheng HE ; Shu HE ; Junhua TANG ; Hong YANG ; Xiaolong LI
Chinese Journal of Radiology 2014;48(7):559-562
Objective To investigate the value of mesenteric vascular CTA in the diagnosis of small intestinal neoplasms.Methods A retrospective analysis of mesenteric CTA from January 2008 to April 2013 was conducted in 51 patients with pathologically proven small intestinal neoplasms.Features of intestinal neoplasms CTA signs including neoplasm feeding artery,draining vein,mesangial side vasa recta and the formed neoplasm vessels,were observed.Two radiologists individually used two methods,namely intestinal tumor feeding artery positioning method and Cole fractionation method,for diagnosis and localization diagnosis of tumor,and also for comparing the results with those of surgical pathology.McNemar Chi-square test was adopted to evaluate the diagnosis differences between the two physicians and between the two methods by the same physician.Kappa value was used to assess the consistency of the results.Results Features of intestinal tumors CTA signs:12 cases of enlarged neoplasm feeding artery,9 cases of early displayed draining vein,22 cases of enlarged mesangial side vasa recta,and 11 cases of vessels formed inside and around the neoplasm,single lesion for all and the largest lesion diameter≥ 5 cm for 37 cases.The accuracy of Cole fractionation method positioning and the feeding artery positioning were 84.31%(43/51) and 98.03%(50/51) respectively.Moderate consistency(Kappa=0.49,P<0.01) was seen with Cole fractionation method by the two physicians and high consistency(Kappa=1.00,P<0.01) with feeding artery positioning method.McNemar Chi-square test showed no significant difference between the two methods by the same physician and the consistency of the results from the two methods was passable(P were 0.062 and 0.125).Conclusion Mesenteric CTA can display the intestinal tumor feeding arteries and draining veins,and is helpful in identification of the relationship between the tumor and its surrounding blood vessels,which can improve the accuracy of pre-operative localization and qualitative diagnosis for small intestinal tumor.
4.Congenital tracheobiliary fistula treated with biological glue plugging under bronchoscopic guidance:a case report and literature review
Xia CHEN ; Yuqun YAN ; Wenhua DENG ; Jiahong REN ; Junhua SHU ; Yabin WU
Chinese Journal of Applied Clinical Pediatrics 2018;33(11):839-841
Objective To investigate the clinical characteristics,diagnosis and treatment of congenital tracheobiliary fistula (CTBF) in children.Methods A case of CTBF admitted into the Department of Pediatric Respiration,Hubei Maternal and Child Health Care Hospital in 2016 was reported,and the related literatures were reviewed.The clinical features,diagnostic methods,treatment status and clinical outcomes of the disease were analyzed.Results The patient was 3 years and 7 months old.The main clinical manifestations were recurrent cough,pneumonia and atelectasis.CTBF was diagnosed by means of iodine oil radiography and treated with biological glue plugging under bronchoscopic guidance.So far,only 30 cases of CTBF have been reported in the English literatures,but only 2 cases in the Chinese literatures.The main clinical manifestations were cough,dyspnea,sputum or bile vomiting,aspiration pneumonia,atelectasis or emphysema.Diagnostic methods for CTBF included bronchoscopy,bronchial angiography and cholangiography,hepatobiliary scan,CT scan and magnetic resonance imaging.Except for this case treated with biological glue plugging under bronchoscopic guidance,all other patients were treated with surgery.The operation methods included fistula ligation,gastrostomy,liver resection,fistula jejunum Roux-en-Y anastomosis,hepatic hilum jejunum anastomosis,gallbladder jejunum anastomosis,etc.Only 4 cases died,and the rest of the patients recovered.Conclusions CTBF should be suspected in children with persistent chronic cough.Iodine oil radiography through bronchoscopy is a simple and feasible method for diagnosis of CTBF.Besides surgery,the lavage and the biological glue plugging method through bronchoscopy is also an effective way to treat CTBF in children without severe biliary malformation.
5.Effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in peripheral blood of patients with psoriasis
Lei MA ; Haibo XUE ; Meilan GAO ; Chunmei SHU ; Juan YU ; Junhua ZHANG
Chinese Journal of Dermatology 2018;51(3):189-193
Objective To evaluate the effect of Notch 1 signaling pathway on the differentiation and function of Th17 cells in peripheral blood of patients with psoriasis.Methods Peripheral blood samples were obtained from 35 patients with psoriasis and 32 healthy controls.Flow cytometry was performed to determine the proportion of Th17 cells in CD4+ T cells in peripheral blood mononuclear cells (PBMCs),real-time RT-PCR to measure the mRNA expression of retinoic acid receptor-related orphan receptor γt (RORγt),interleukin (IL)-17,Notch 1 and hairy-and-enhancer-of-split-1 (Hes-1),and enzymelinked immunosorbent assay (ELISA) to detect levels of IL-17 in the serum and culture supernatant of PBMCs stimulated with phorbol ester,calcium ionophore and brefeldin A.The correlation of Notch 1 mRNA expression with psoriasis area and severity index (PASI) was analyzed,so were its correlation with the proportion of Th17 cells,mRNA expression of RORγt,and mRNA and protein expression of IL-17.PBMCs isolated from the patients with psoriasis were divided into 5 groups to be treated with γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t-butyl ester (DAPT) at different concentrations of 0(control group),2.5,5.0,10.0 and 20.0μmol/L,respectively,so as to evaluate the effects of blocking the Notch1 signaling pathway by DAPT on the proportion of Th17 cells in PBMCs,levels of RORγt and IL-17.Results Compared with the healthy controls,patients with psoriasis showed a significant increase in the proportion of Th17 cells in CD4+ T cells in PBMCs (2.863% ± 0.969% vs.0.604% ± 0.124%,P < 0.01),mRNA expression of RORγt (5.255 ± 0.998 vs.1.530-± 0.485,P < 0.01),Notch1 (6.743 ± 1.756 vs.1.731 ± 0.456,P < 0.01),Hes-1 (6.384 ± 1.665 vs.1.627 ± 0.485,P < 0.01) and IL-17 (6.944 ± 1.626 vs.1.698 ± 0.329,P < 0.01),and serum level of IL-17 ([36.444 ± 5.936] ng/L vs.[11.762 ± 2.260] ng/L,P < 0.01).Among the patients with psoriasis,the mRNA expression of Notch1 was positively correlated with PASI scores,proportion of Th17 cells,mRNA expression of RORγt and IL-17,and serum level of IL-17 (r =0.584,0.544,0.518,0.549 and 0.511,respectively,all P < 0.05).There were significant differences in the proportion of Th17 cells,mRNA expression of RORγt and IL-17,and level of IL-17 in the culture supematant among the control group,2.5-,5.0-,10.0-and 20.0-μmol/L DAPT groups (F =79.527,82.239,78.086 and 80.558,respectively,all P < 0.01).The above indices were significantly lower in the 2.5-,5.0-,10.0-and 20.0-μmol/L DAPT groups than in the control group (all P < 0.05),and decreased along with the increase of DAPT concentrations.Conclusion Notch1 signaling pathway can promote the differentiation of Th17 cells and the expression of RORγt,IL-17 and Hes-1 in the peripheral blood of patients with psoriasis.
6.Clinical value of nucleic acid detection for hepatitis B virus screening in hospitalized patients
Chunhong DU ; Junhua HU ; Yuan ZHANG ; Jiwu GONG ; Jun ZHOU ; Qin MENG ; Juan LIU ; Jiangcun YANG ; Rong GUI ; Xianping LYU ; Rong XIA ; Fenghua LIU ; Li QIN ; Shu SU ; Jinqi MA ; Juan CAI ; Huifang JIN ; Qi ZHANG ; Jun ZHANG ; Rongyi CAO ; Xiying LI ; Peng WANG
Chinese Journal of Laboratory Medicine 2023;46(1):27-31
Objective:To explore clinical value of nucleic acid detection for hepatitis B virus (HBV) screening in hospitalized patients.Methods:This cross-sectional study collected and analyzed plasma samples from patients admitted to 10 domestic medical institutions from July 2021 to December 2021. Serological immunoassay and nucleic acid screening were used to simultaneously detect hepatitis B markers such as hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B e Antigen (HBeAg), hepatitis B e antibody (HBeAb), hepatitis B core antibody (HBcAb),and HBV DNA. Statistical analysis was performed on the serology, nucleic acid test results and clinical information of the patients.Results:Of the 8 655 collected samples, HBsAg was positive in 216 (2.50%) samples,HBV DNA was positive in 238 (2.75%) samples ( P>0.05); 210 (2.43%) samples were positive for both HBsAg and HBV DNA, 28 (0.32%) were HBsAg negative and HBV DNA positive, 6 cases (0.07%) were HBsAg positive and HBV DNA negative. Conclusion:These results indicate that the HBV DNA testing is equally effective as hepatitis B virus serological detection for hepatitis B virus screening in hospitalized patients.
7.Cost-effectiveness analysis of nucleic acid screening for hepatitis B and C in hospitalized patients in China
Shu SU ; Qi ZHANG ; Peng WANG ; Rong GUI ; Chunhong DU ; Xiying LI ; Xianping LYU ; Rong XIA ; Fenghua LIU ; Li QIN ; Jiameng NIU ; Lili XING ; Leilei ZHANG ; Jinqi MA ; Junhua HU ; Yuan ZHANG ; Juan CAI ; Huifang JIN ; Jun ZHANG ; Rongyi CAO ; Jiwu GONG ; Jiangcun YANG
Chinese Journal of Laboratory Medicine 2023;46(1):38-44
Objective:To compare the cost-effectiveness of hospitalized Chinese patients undergoing nucleic acid screening strategies for hepatitis B and hepatitis C, immunological screening strategy, and no screening strategy under different willingness to pay (WTP). The results might aid to decision-making for the optimal strategy.Methods:In this study, nucleic acid screening, immunological screening and no screening were used as screening strategies, and China′s GDP in 2021 (80 976 yuan) was used as the threshold of WTP to construct a Markov model. After introducing parameters related to the diagnosis and treatment of hepatitis B and C in inpatients, a cohort population of 100 000 inpatients was simulated by TreeAge Pro 2021 software, the total cost, total health effects, incremental cost-effectiveness ratio and average cost-effectiveness ratio of different screening strategies were calculated, and cost-effectiveness analysis was conducted. Univariate and probabilistic sensitivity analysis were used to assess the impact of parameter uncertainty on the final results.Results:Compared with the non-screening strategy, the incremental total cost of the hepatitis B immunological screening strategy for cohort patients was 11 049 536 yuan, and the incremental cost-effectiveness ratio was 24 762 yuan/quality-adjusted life years (QALY), while the total incremental cost of nucleic acid screening was 19 208 059 yuan, and the incremental cost-effectiveness ratio was 29 873 yuan/QALY; the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 834 yuan/QALY. Compared with the non-screening strategy, the incremental cost-effectiveness ratio of hepatitis C immunological screening strategy was 5 731 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening strategy was 8 722 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 591 yuan/QALY. The results of probabilistic sensitivity analysis showed that when the cost of nucleic acid testing exceeded 214.53 yuan, it was not cost-effective to perform hepatitis B nucleic acid screening under the WTP as 1 fold GDP. When the cost of nucleic acid testing exceeded 132.18 yuan, it was not cost-effective to conduct hepatitis C screening under the WTP as 1 fold GDP.Conclusions:Nucleic acid screening strategy can achieve more cost-effectiveness and is worthy of vigorous promotion. Compared with no screening, both the nucleic acid and immunological screening strategies are cost-effective, and hepatitis nucleic acid screening is the optimal strategy for hospitalized patients.
8.A multicenter study assessing the efficacy of various preoperative/pre-transfusion screening methods for blood transmitted disease
Junhua HU ; Li QIN ; Juan LIU ; Xinghuan MA ; Qin MENG ; Peng WANG ; Jiangcun YANG ; Rong GUI ; Chunhong DU ; Xiying LI ; Xianping LYU ; Rong XIA ; Fenghua LIU ; Shu SU ; Jinqi MA ; Yuan ZHANG ; Juan CAI ; Huifang JIN ; Qi ZHANG ; Jun ZHANG ; Rongyi CAO ; Bing HAN ; Jiwu GONG ; Jun ZHOU
Chinese Journal of Laboratory Medicine 2023;46(1):32-37
Objective:This multi-centre study was conducted to assess the efficacy of various preoperative/pre-transfusion screening methods for blood transmitted disease.Methods:From July 2021 to December 2021, plasma samples of patients admitted to 10 hospitals were collected for screening preoperative/pre-transfusion blood transmitted disease. Nucleic acid detection technology was used to detect hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA and human immunodeficiency virus (HIV)(1+2) RNA, and the results were compared with the immuno-serological methods. χ 2 test and Kappa test were used to analyze the efficacy of these two methods. Results:A total of 8 655 valid specimens were collected from 10 hospitals. There was a statistically significant difference in the positive detection rate of HCV between the two methods ( P<0.001). There was no significant difference in the positive detection rate of HBV and HIV assessed by the two methods ( P>0.05), but the number of positive cases detected by HBV DNA and HIV RNA (218 and 4 cases) was significantly higher than the corresponding serological results (216 and 2 cases). At the same time, there were HBV, HCV and HIV immuno-serological omissions by the immuno-serological methods, among which 28 cases were HBsAg negative and HBV DNA positive, 2 cases were HCV antibody negative and HCV RNA positive, and 2 cases were HIV antigen/antibody negative and HIV RNA positive. In addition, in the 66 samples with inconsistent results from the two detection methods, 83.3% (55/66), 68.2% (45/66), 63.6% (42/66) and 62.1% (41/66) of patients aged was>45 years, tumor, surgery and male, respectively. Conclusions:Compared with immuno-serological tests, nucleic acid tests have the advantage in terms of sensitivity on detecting HBV, HCV and HIV infection and could reduce missed detection. The risk of transmission can be reduced by adding HBV, HCV, and HIV nucleic acid tests to preoperative/pre-transfusion immuno-serological tests screening for patients over 45 years of age and tumor patients.