Serum immunoglobulin concentration and the risk of type 2 diabetes mellitus in adults in Tianjin City: a prospective cohort study / 中华预防医学杂志

Objective: To evaluate the association between immunoglobulin concentration and the risk of type 2 diabetes mellitus (T2DM) in adults in Tianjin City. Methods: Based on the Tianjin Chronic Low-grade Systemic Inflammation and Health (TCLSIHealth) cohort from January 2010 to December 2018, subjects who had completed the measurement of baseline immunoglobulin concentration and blood glucose concentration and not been diagnosed with any type of diabetes at baseline were selected in this study. The collected data included the concentration of serum immunoglobulin (IgG, IgM, IgA and IgE), fasting blood glucose and other potential confounders. The subjects were divided into four groups from Q1 to Q4 according to the quartiles of baseline immunoglobulin concentration. The multivariable Cox regression model was used to assess the association between the baseline immunoglobulin concentration and T2DM. Results: A total of 6 315 subjects aged (50.1±10.0) years were included. About 390 subjects were newly diagnosed with T2DM during the follow-up period. The incidence rate was 16.8/1 000 person-years. After adjusting for age, sex, waist circumference, smoking status, drinking status, eosinophil ratio, metabolic syndrome, first-or second-degree family history, and reciprocal adjusting for other immunoglobulin concentrations, compared to the lowest quartile concentration group Q1, subjects in group Q4 with the highest quartile of IgG concentration showed a lower risk of T2DM (HR=0.71, 95%CI: 0.52-0.97), and subjects in group Q4 with the highest quartile of IgM concentration also had a decreased risk of T2DM (HR=0.66, 95%CI: 0.47-0.91). Subjects in group Q4 with the highest quartile of IgA concentration had an increased risk of T2DM (HR=1.56, 95%CI: 1.18-2.07). The risk of T2DM decreased with the increase of serum IgG and IgM concentrations (Ptrend=0.018, Ptrend=0.010) and increased with the increase of serum IgA concentrations (Ptrend<0.001). No association was found between the concentration of IgE and T2DM risk (HR=0.99, 95%CI: 0.74-1.31, Ptrend=0.891). Conclusion: The concentration of IgG and IgM is negatively associated with the risk of T2DM, and the concentration of IgA is positively associated with the risk of T2DM in Tianjin City. The concentrations of IgG, IgM and IgA could be a predictor of hyperglycemia and T2DM.

Serum immunoglobulin concentration and the risk of type 2 diabetes mellitus in adults in Tianjin City: a prospective cohort study / 中华预防医学杂志

Objective: To evaluate the association between immunoglobulin concentration and the risk of type 2 diabetes mellitus (T2DM) in adults in Tianjin City. Methods: Based on the Tianjin Chronic Low-grade Systemic Inflammation and Health (TCLSIHealth) cohort from January 2010 to December 2018, subjects who had completed the measurement of baseline immunoglobulin concentration and blood glucose concentration and not been diagnosed with any type of diabetes at baseline were selected in this study. The collected data included the concentration of serum immunoglobulin (IgG, IgM, IgA and IgE), fasting blood glucose and other potential confounders. The subjects were divided into four groups from Q1 to Q4 according to the quartiles of baseline immunoglobulin concentration. The multivariable Cox regression model was used to assess the association between the baseline immunoglobulin concentration and T2DM. Results: A total of 6 315 subjects aged (50.1±10.0) years were included. About 390 subjects were newly diagnosed with T2DM during the follow-up period. The incidence rate was 16.8/1 000 person-years. After adjusting for age, sex, waist circumference, smoking status, drinking status, eosinophil ratio, metabolic syndrome, first-or second-degree family history, and reciprocal adjusting for other immunoglobulin concentrations, compared to the lowest quartile concentration group Q1, subjects in group Q4 with the highest quartile of IgG concentration showed a lower risk of T2DM (HR=0.71, 95%CI: 0.52-0.97), and subjects in group Q4 with the highest quartile of IgM concentration also had a decreased risk of T2DM (HR=0.66, 95%CI: 0.47-0.91). Subjects in group Q4 with the highest quartile of IgA concentration had an increased risk of T2DM (HR=1.56, 95%CI: 1.18-2.07). The risk of T2DM decreased with the increase of serum IgG and IgM concentrations (Ptrend=0.018, Ptrend=0.010) and increased with the increase of serum IgA concentrations (Ptrend<0.001). No association was found between the concentration of IgE and T2DM risk (HR=0.99, 95%CI: 0.74-1.31, Ptrend=0.891). Conclusion: The concentration of IgG and IgM is negatively associated with the risk of T2DM, and the concentration of IgA is positively associated with the risk of T2DM in Tianjin City. The concentrations of IgG, IgM and IgA could be a predictor of hyperglycemia and T2DM.

Celastrol induced hepatocellular carcinoma cell death via ROS-mediated DNA damage / 中国药理学通报

Aim To investigate the role of eelastrol in reactive oxygen species ( ROS) accumulation and DNA damage in hepatocellular carcinoma cells, and further investigate its effect on apoptosis induction in cancer cells.Methods Human liver cancer HepG2 and Huh7 cells were cultured with celastrol, then the morphological changes of cells were observed under microscope.MTT assay was employed to detect the proliferation of hepatocellular carcinoma cells, CM-H2DCFDA probe to detect intracellular ROS levels, and immunofluorescence to detect the expression level of -y-H2AX in celastrol-treated cells.Hoechst 33258 staining was used to observe nuclear condensation and fragmentation; Flow cytometry was used to evaluate cell death.J J Western blot was applied to measure the expression levels of -y-H2 AX, caspase-3, PARP and other proteins.Results Celastrol had a significant inhibitory effect on the proliferation of liver cancer cells in dose-dependent manner.Comparer] with the eontrol group, the eell viability was reduced and intracellular ROS level also increased significantly after celastrol treatment in a dose-dependent manner ( P < 0.05 ).With Hoechst staining, typical apoptotic characteristics such as nuclear chromatin condensation and fragmentation were observed in celastrol-treated cells.Western blot results showed that pro-form of caspase-3 significantly decreased, and the cleavage of PARP markedly increased by celastrol.After pretreatment with ROS inhibitor NAC, celastrol-mediated caspase-3 activation and PARP cleavage were significantly reversed ( P < 0.05 ).Conclusions Celastrol can induce apoptosis in hepatocellular carcinoma cells, and its anti-cancer effect is dependent on ROS-mediated DNA damage.

Clinical and pathological features in IgA nephropathy with IgG deposition in the glomerular mesangial area / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relationship between the clinical and pathological findings in IgA nephropathy with or without IgG deposition in the glomerular mesangial area.</p><p><b>METHODS</b>The data were collected from 122 patients with a diagnosis of IgA nephropathy by renal biopsy in the Third Affiliated Hospital of Southern Medical University between November, 2009 and February, 2016. All the samples were examined by light microscopy, immunofluorescence and electron microscopy. According to the results of immunofluorescence assay, the patients were divided into IgA group (n=63) and IgA-IgG group (n=59). The pathological classification of IgA nephropathy was analyzed according to Oxford classification and Lee's classification. The clinical and pathological findings were compared between the two groups.</p><p><b>RESULTS</b>Compared with the patients with IgA nephropathy but without IgG deposition, patients with IgA nephropathy with IgG deposition had higher serum creatinine, higher 24-h urine protein, higher blood uric acid, higher triglyceride levels (P<0.05) and lower eGFR (P<0.05); more of these patients were in Lee's grade IV-V, had renal tubular atrophy and/or interstitial fibrosis, and had MEST scores more than 3 (P<0.05).</p><p><b>CONCLUSION</b>Patients with IgA nephropathy with IgG deposition in the glomerular mesangial have severer clinical symptoms and more serious pathological changes. Measures should be taken to control IgG deposition in patients with IgA nephropathy to delay the progress of the disease.</p>

Association of waist-to-hip ratio with chronic kidney disease in non-diabetic subjects / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between waist-to-hip ratio (WHR) and chronic kidney disease (CKD) in non-diabetic subjects and compare the difference between male and female subjects.</p><p><b>METHODS</b>We performed a cross-sectional survey among 2142 community-based southern Chinese participants without diabetes from June to October 2012. We divided all the participants into 4 groups according to the gender-specific quartiles of WHR. Logistic regression models were used to explore the associations of WHR with CKD in these subjects.</p><p><b>RESULTS</b>In the unadjusted model, WHR was significantly associated with CKD in women (OR=7.29, 95% CI: 3.56-16.32, P<0.001), and the association was still significant (OR=6.13, 95% CI: 2.56-15.20, P=0.003 ) after adjustment for the potential confounders (including age, history of hypertension, coronary heart disease, current smoker, physical inactivity, education level, systolic blood pressure, diastolic blood pressure, serum triglyceride, serum high density lipoprotein, blood glucose, and BMI). The odds ratio (OR) for having CKD in the highest versus lowest quartile of WHR levels was 2.44 (95% CI: 0.98-4.97, P=0.103) in men in the unadjusted model.</p><p><b>CONCLUSION</b>WHR levels are associated with CKD in non-diabetic women but not in non-diabetic male subjects.</p>

Prevalence and distribution of metabolic syndrome in a southern Chinese population Relation to exercise, smoking, and educational level

To investigate the prevalence and distribution of metabolic syndrome [MetS] and the impact of exercise, smoking, and educational level on the risk of MetS in a southern Chinese population. A cross-sectional study was conducted in Zhuhai City, China from June to August 2012. Data on exercise, smoking, and educational level, anthropometric parameters, blood pressure, lipid, and glucose levels were collected. The prevalence of MetS [as defined by the International Diabetes Federation] was determined. Data necessary to evaluate MetS, the socio-economic characteristics, and lifestyle were obtained for 4645 subjects aged 18-75 years old. A total of 19.8% of the participants had MetS. The adjusted odds of having MetS were lower among males [adjusted odds: 0.75; 95% confidence interval [CI]: 0.57-1.01] compared with females. Those participants who currently smoked had a higher risk of developing MetS compared with non-smokers [adjusted odds: 1.61; 95% CI: 1.13-2.50]. Those who had no physical exercise had a higher risk of developing MetS compared with those who physically exercised more than 60 minutes/day [adjusted odds: 1.51; 95% CI: 1.12-2.23;]. Compared with those with no education, every category of attained educational level had a lower risk of developing MetS [P<0.001]. The findings in this study revealed that current smokers had a greater risk of developing MetS compared with non-smokers. Increased physical activity and higher levels of education attained served as protective factors for the population

Preliminary study on the quantity and function of T follicular helper cells in the cytopenic patients with positive BMMNC-Coombs test / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the quantity and function of bone marrow (BM) T follicular helper (Tfh) cells of the cytopenia patients with positive bone marrow mononuclear cells (BMMNC)- Coombs test (also known as immuno-related pancytopenia, IRP), and explore the role of Tfh cells in the pathogenesis of IRP.</p><p><b>METHODS</b>Forty- three untreated IRP patients, 47 recovered IRP patients and 25 healthy donors were enrolled in this study. The percentages of Tfh cells, Tfh-related molecules ICOS, CD40L, IL-21 and Bcl-6 in BM were investigated by flow cytometry and semiquantitive RT-PCR.</p><p><b>RESULTS</b>The ratio of CD4⁺CXCR5⁺/CD4⁺ cells of untreated IRP patients [(28.79 ± 19.70)%] was significantly higher than that of recovered IRP patients [(21.15 ± 12.81)% ] and normal controls ([ 13.42 ± 6.72)% ](P<0.05). The ratio of CD4⁺CXCR5⁺ICOS⁺/CD4⁺CXCR5⁺ cells of untreated IRP patients [(5.05 ± 4.71)% ] was significantly higher than that of recovered IRP patients [(2.96 ± 2.89)% ] and normal controls [(2.99 ± 2.23)% ] (P<0.05). The ratio of CD4⁺CXCR5⁺CD40L⁺/CD4⁺CXCR5⁺ cells of untreated IRP patients [(5.87 ± 4.14)%] and recovered IRP patients [(6.52±5.47)%] were significantly higher than that of normal controls [(2.93 ± 2.92)%] (P<0.05). The ratio of intracytoplasmic CD4⁺CXCR5⁺IL-21⁺/CD4⁺CXCR5⁺ cells of untreated IRP patients [(8.20 ± 7.41)% ] and recovered IRP patients [(6.30 ± 6.03)% ] were significantly higher than that of normal controls [(3.43 ± 3.40)%] (P<0.05). The relative expressions of Bcl-6 mRNA in BMMNC were 0.625 ± 0.248, 0.485 ± 0.253, 0.306 ± 0.210 in three groups, respectively. The differences between untreated IRP patients, recovered IRP patients and normal controls were significant (P<0.05).</p><p><b>CONCLUSION</b>There exists increased quantity and hyperfunction of Tfh cells in the IRP patients, they may play important role in the pathogenesis of IRP. Tfh cells and their related effector molecules could be a potential therapeutic target for the disease.</p>

The mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test (IRP).</p><p><b>METHODS</b>Twenty-one iron overloading, 26 non-iron overloading IRP patients and 10 normal controls were enrolled in this study. The expressions of ROS, Bcl-2, Caspase-3 and apoptosis of BMMNC were analyzed by flow cytometry (FCM). Antioxidants were added to iron overloading IRP BMMNC, and then the changes of indices above were detected by FCM. The number and apoptosis of T lymphocytes of IRP patients were also detected.</p><p><b>RESULTS</b>ROS and apoptosis of BMMNC, myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones and normal controls (P < 0.05). The expressions of Bcl-2 on BMMNC, erythrocytes and stem cells of iron overloading IRP patients were significantly lower than those of non-iron overloading IRP ones (P < 0.05). The levels of Caspase-3 on myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones and normal controls (P < 0.05). After treatment with antioxidants, the expressions of ROS, Caspase-3 and apoptosis of iron overloading IRP BMMNC significantly decreased, but opposite for Bcl-2. The percentages of CD4(+) lymphocytes [ ( 40.86 ± 8.74）%] and CD4(+)/CD8(+) (1.44 ± 0.36) in PB of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones [(35.96 ± 7.03)% and 1.14 ± 0.37] and normal controls [(28.00 ± 6.73)% and 0.79 ± 0.21], respectively (P < 0.05), as opposite for CD8(+) lymphocytes (P < 0.05). The apoptosis of CD8(+) lymphocytes [(27.35 ± 10.76)%] and the ratio of CD8(+) apoptosis/CD4(+) apoptosis (2.51 ± 0.81) in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones [(15.47 ± 8.99)%] and normal controls (1.39 ± 0.47), respectively (P < 0.05). The apoptosis of erythrocytes and stem cells coated with auto-antibodies in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP and normal controls.</p><p><b>CONCLUSION</b>Mechanisms underlying bone marrow damage by iron overload might be through the follows: ①The increased ROS induced by excessive iron deposition affected the expressions of Caspase-3 and Bcl-2, which caused more BMMNC apoptosis; ②The abnormal number and ratio of T lymphocytes caused by iron overload aggravated the abnormality of immunity of IRP; ③Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.</p>

Effects of docosahexaenoic acid on ion channels of rat coronary artery smooth muscle cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of docosahexaenoic acid (DHA) on large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels and voltage-dependent K(+) (K(V)) channels in rat coronary artery smooth muscle cells (CASMCs), and evaluate the vasorelaxation mechanisms of DHA.</p><p><b>METHODS</b>BK(Ca) and K(V) currents in individual CASMC were recorded by patch-clamp technique in whole-cell configuration. Effects of DHA at various concentrations (0, 10, 20, 40, 60 and 80 µmol/L) on BK(Ca) and K(V) channels were observed.</p><p><b>RESULTS</b>(1) DHA enhanced IBK(Ca) and BK(Ca) tail currents in a concentration-dependent manner while did not affect the stably activated curves of IBK(Ca). IBK(Ca) current densities were (68.2 ± 22.8), (72.4 ± 24.5), (120.4 ± 37.9), (237.5 ± 53.2), (323.6 ± 74.8) and (370.6 ± 88.2)pA/pF respectively (P < 0.05, n = 30) with the addition of 0, 10, 20, 40, 60 and 80 µmol/L DHA concentration, and half-effect concentration (EC(50)) of DHA was (36.22 ± 2.17)µmol/L. (2) IK(V) and K(V) tail currents were gradually reduced, stably activated curves of IK(V) were shift to the right, and stably inactivated curves were shifted to the left in the presence of DHA. IK(V) current densities were (43.9 ± 2.3), (43.8 ± 2.3), (42.9 ± 2.0), (32.3 ± 1.9), (11.7 ± 1.5) and (9.6 ± 1.2)pA/pF respectively(P < 0.05, n = 30)post treatment with 0, 10, 20, 40, 60 and 80 µmol/L DHA under manding potential equal to +50 mV, and EC(50) of DHA was (44.19 ± 0.63)µmol/L.</p><p><b>CONCLUSION</b>DHA can activate BK(Ca) channels and block K(V) channels in rat CASMCs, the combined effects on BK(Ca) and K(V) channels lead to the vasodilation effects of DHA on vascular smooth muscle cells.</p>

Quantity and subtypes of dendritic cells in patients with immune related pancytopenia and their clinical significance / 中国实验血液学杂志

This study was aimed to investigate the quantity and subtypes of dendritic cells (DC) in patients with immune related pancytopenia (IRP) and to explore the role of DC in pathogenesis of IRP. The quantity of plasmacytoid dendritic cells (pDC, Lin(-)HLA-DR(+) CD123(+) cells) and myeloid dendritic cells (mDC, Lin(-)HLA-DR(+) CD11c(+)cells) in peripheral blood of 65 patients with IRP (37 new diagnosed and 28 remitted) and 17 healthy controls were analyzed by flow cytometry. The results indicated that the ratio of pDC in peripheral blood mononuclear cells (PBMNC) was (0.91 ± 064)% in new diagnosed group, which was significantly higher than that in remission group (0.39 ± 0.11)% and control group (0.29 ± 0.13)% (P < 0.01), while this ratio of pDC in remission group was higher than that in control group (P < 0.05). The ratio of mDC in PBMNC was (0.21 ± 0.20)% in new diagnosed group and (0.34 ± 0.21)% in remission group respectively, there was no statistical difference as compared with control group (0.29 ± 0.09)% (P > 0.05). The ratio of pDC to mDC in new diagnosed group was 6.75 ± 7.11, which was significantly higher than that in remission group (1.55 ± 0.93) and control group (1.07 ± 0.43, P < 0.01), there was no statistical difference between the ratio of remission group and control group (P > 0.05). The ratio of pDC in PBMNC of IRP group negatively correlated to ratio of Th1/Th2 (r = -0.347, P < 0.05), and positively correlated to the ratio of auto-antibody on membrane of BMMNC (r = 0.606, P < 0.05) and to the quantity of CD5(+)B cells (r = 0.709, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.381, P < 0.01) and platelets (r = -0.343, P < 0.01). The ratio of mDC in PBMNC positively correlated to the ratio of Th1/Th2 (r = 0.595, P < 0.05) and the level of hemoglobin (r = 0.292, P < 0.05). The ratio of pDC/mDC negatively correlated to ratio of Th1/Th2 (r = -0.395, P < 0.05), it positively correlated to the level of antibody on membrane of BMMNC (r = 0.421, P < 0.05) and the quantity of CD5(+)B cells (r = 0.423, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.304, P < 0.05) and platelets (r = -0.287, P < 0.05). It is concluded that the quantity of pDC in peripheral blood of IRP patients increases, which may be related to the immunopathogenesis of IRP.

Application of bone marrow indirect Coombs test and its clinical significance in diagnosis of immuno-related pancytopenia / 中国实验血液学杂志

This study was purposed to applicate the bone marrow indirect Coombs test and investigate its clinical significancies in diagnosis of immuno-related pancytopenia (IRP). 30 patients with pancytopenia including 22 cases of IRP and 8 cases of idiopathic cytopenia of undetermined significance (ICUS), and 15 patients with iron-deficiency anemia as controls were enrolled in this study. After incubation of the bone marrow supernatant of IRP patients and bone marrow nucleated cell (BMNC) of controls was used as experiment group, while the incubation of BMNC and bone marrow supernatant of controls was used as control group. After incubation for 45 min, the positive rate of membrane antibodies in bone marrow hematopoietic cells (CD15(+), GlyCoA(+) and CD34(+)cells) was detected by flow cytometry, and correlation analysis of positive rate with clinical data of patients were analyzed. The results showed that among 30 patients with pancytopenia (16 positive and 14 negative for bone marrow direct Coombs test) 16 cases showed positive for bone marrow indirect Coombs test, with positive rate 53.33. In the experiment group, the median positive rate of CD15(+)IgM was 0.34, which was significantly higher than that in control group (0.20, P < 0.05); the median positive rates of CD34(+) IgG and IgM were 0.64 and 0.21 respectively, which were significantly higher than those in control group (0.00, P < 0.05) and (0.00, P < 0.05); the positive rates of GlyCoA(+)IgG and IgM were (0.83 ± 0.75) and (2.12 ± 1.98) respectively, which were significantly higher than those in control group [(0.47 ± 0.43), P < 0.05, (0.68 ± 0.64), P < 0.01]; the positive rates of CD15(+) IgG and IgM were positively correlated with the ratio of CD5(+)B cells. The positive rates of GlyCoA(+) IgG and IgM negatively correlated with the Hb level, percentage of reticulocytes, the ratio of bone marrow erythroid lineage and DC1/DC2 positively correlated with the ratio of CD5(+)B cells and indirect bilirubin level. It is concluded that antibodies (IgG or IgM) aiming at the bone marrow hematopoietic cells exist in the supernatant of some IRP and ICUS patients, and may act on the membrane protein of the normal BMNC. These antibodies correlate with the prognosis of IRP.

Expression of CD80 and CD86 on dendritic cells of patients with immune related pancytopenia and its clinical significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the function of dendritic cells (DC) of patients with immune related pancytopenia (IRP) and explore the role of DC in IRP.</p><p><b>METHODS</b>The expression of CD80 and CD86 on myeloid DC (mDC, Lin-HLA-DR(+) CD11c(+) cells) and plasmacytoid DC (pDC, Lin-HLA-DR(+) CD123(+) cells) of 65 IRP (37 untreated and 28 remitted) patients and 17 healthy controls were analyzed by flow cytometry.</p><p><b>RESULTS</b>The expression of CD86 on pDC was (82.47 ± 13.17)% in untreated group and (60.08 ± 14.29)% in remission group, which were significantly higher than that of controls (47.95 ± 18.59)% (P < 0.05), while the expression in untreated group was higher than that of remission group (P < 0.05). The expression of CD80 on pDC was (6.31 ± 4.49)% in untreated group, which was significantly higher than that of remitted patients (3.09 ± 2.93)% and controls (2.33 ± 2.25)% (P < 0.05). The expression of CD86 on mDC was (97.06 ± 4.82)% in untreated group and (91.35 ± 12.20)% in control group, while the expression in untreated group was higher than that of control group (P < 0.05). The expression of CD80 on mDC was (6.20 ± 5.44)% in untreated group and (3.97 ± 3.24)% in remission group, which were significantly higher than that of controls (1.86 ± 1.73)% (P < 0.05). The expression of CD86 on pDC was negatively correlated to Th1/Th2 (r = -0.733, P < 0.05), it was positively correlated to the antibody on membrane of BMMNC (r = 0.283, P < 0.05) and the quantity of CD5(+)B cells (r = 0.436, P < 0.05), while it was negatively correlated to the level of hemoglobin, platelets and white blood cells (r = -0.539, P < 0.05; r = -0.519, P < 0.05; r = -0.567, P < 0.05, respectively). The expression of CD80 on pDC was negatively correlated to the level of hemoglobin and platelets (r = -0.431, P < 0.05; r = -0.464, P < 0.05).</p><p><b>CONCLUSION</b>The function of pDC in PB of IRP were strengthened, which was relevant to the immunopathogenesis of IRP.</p>

EPOR and TPOR expressions on CD34+ CD59- and CD34+ CD59+ bone marrow cells from patients with paroxysmal nocturnal hemoglobinuria / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the expressions of erythropoietin receptor (EPOR) and thrombopoietin receptor (TPOR) on CD34+ CD59- and CD34+ CD59+ bone marrow (BM) cells from patients with paroxysmal nocturnal hemoglobinuria (PNH).</p><p><b>METHODS</b>(1) The expressions of EPOR and TPOR on CD34+ CD59- and CD34+ CD59- BM cells from 26 PNH patients and 16 normal controls were examined by flow cytometry (FCM). (2) The mRNA expression of the EPOR and the TPOR in BM mononuclear cells (BMMNC) from 25 PNH patients and 13 normal controls were examined by RT-PCR.</p><p><b>RESULTS</b>(1) The percentage of EPOR positive cells in PNH CD34+ CD59+ BMMNC [(30.67 +/- 18.30)%] was significantly higher than that in PNH CD34+ CD59- BMMNC [(8.05 +/- 3.51)%] (P < 0.01) and than that in control CD34+ CD59+ BMMNC [(8.24 +/- 6.51)%] (P < 0.01), but there was no obvious difference between the CD34+ CD59-BMMNC in PNH and CD34+ CD59+ BMMNC in control. (2) The percentage of TPOR positive cells in PNH CD34+ CD59+ BMMNC [(28.15 +/- 17.75)%] was significantly higher than that in PNH CD34+ CD59-BMMNC [(15.65 +/- 14.45)%] (P < 0.05) and than that in control CD34+ CD59+ BMMNC [(10.77 +/- .39)%] (P < 0.01), but there was no obvious difference between the CD34+ CD59- BMMNC in PNH and CD34+ CD59+ BMMNC in control. (3) There was no statistic difference in EPOR mRNA and TPOR mRNA expressions in BMMNCs between PNH patients group [(0.41 +/- 0.37) and (0.32 +/- 0.19), respectively] and control group [(0.47 +/- 0.33) and (0.40 +/- 0.29), respectively].</p><p><b>CONCLUSION</b>The expression of EPOR and TPOR of PNH patients on BM CD34+ CD59+ cells are significantly higher than those on BM CD34+ CD59- cells. The difference may be due to abnormal transcription of both receptor coding genes.</p>

Effect of CD8+ effector T cells on the hematopoiesis pathway damage in the patients with severe aplastic anemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the quantity and the pathway to damage hematopoietic cells of CD8+CD25+ and CD8+ HLA-DR+ effector T cells in peripheral blood (PB) of severe aplastic anemia(SAA) patients and explore the immunopathogenesis of SAA.</p><p><b>METHODS</b>The quantity of CD8+ CD25+ and CD8+ HLA-DR+ cells in PB and the expressions of perforin, granzyme B, tumor necrosis factor-beta (TNF-beta) and FasL in 29 SAA (14 untreated and 15 recovered) patients and 12 normal controls were analyzed by flow cytometry.</p><p><b>RESULTS</b>The fraction of CD8+ CD25+ T cells in CD8+ T cells was (3.67 +/- 2.58)% in untreated SAA patients, (5.19 +/- 4. 29)% in recovered patients and (4.84 +/- 2.31)% in normal controls, and that of CD8+ CD25+ T cells in CD3+ cells in the three groups was (2.25 +/- 1.35)%, (2.98 +/- 1.35)% and (2.11 +/- 1.88)%, respectively. They had no statistic difference among the 3 groups (P >0.05). The fraction of CD8+ HLA-DR+ T cells in CD8+ T cells was (39.30 +/- 8.13)% in untreated patients, which was significantly higher than that in recovered patients [(20.65 +/- 5.38)%] and controls [(18.34 +/- 6.68)%] (P<0.001), while there was no statistic difference between the latter two groups (P>0.05). CD8+ HLA-DR+ T cells in CD3+ cells was (27.81 +/- 7.10)% in untreated group, which was significantly higher than that of recovered group [(12.02 +/- 3.03)%] and controls [(8.50 +/-2.33)%] (P<0.01). And that in recovered group was higher than that in control group (P<0.05). The expressions of perforin, granzyme B, TNF-beta and FasL of CD8+ HLA-DR+ T cells in untreated group were 8.51%, 96.08%, 72.11% and 94.25% respectively, which were higher than those in recovered group (1.78%, 85.20%, 34.38% and 51.20%) and controls (1.86%, 82.09% ,17.92% and 32.91%). There was no statistic difference between recovered patients and controls (P>0.05).</p><p><b>CONCLUSION</b>There were elevated quantity of CD8+ HLA-DR+ T cells and high expressions of perforin, granzyme B, TNF-beta and FasL in SAA, which might contribute to the bone marrow failure.</p>

Quantity and function of T cell subsets in patients with paroxysmal nocturnal hemoglobinuria / 中国实验血液学杂志

This study was purposed to investigate the immune state of T cells, the quantity and function of GPI(+) T cells and GPI(-) T cells in patients with paroxysmal nocturnal hemoglobinuria (PNH). 22 cases of PNH and 18 normal controls were enrolled in this study. Their T lymphocyte subsets, Th lymphocyte subsets were assayed by flow cytometry with the monoclonal antibodies concerned. The proportion of GPI(+) T cells or GPI(-) T cells in CD3(+) T cells, CD4(+) T cells, CD8(+) T cells and the expressions of CD69 on these T cells were also respectively assayed. The results showed that the proportion of CD4(+) T cells in CD3(+) T cells in PNH [(47.7670 +/- 13.91139)%] was lower than that in controls [(54.9592 +/- 7.11678)%] (p < 0.05). CD8(+) T cells in CD3(+) T cells of PNH cases [(52.2767 +/- 13.90395)%] were higher than that of controls [(45.2418 +/- 6.75306)%] (p < 0.05). The ratio of CD4(+) T cells to CD8(+) T cells was reverse in PNH. Those were more significantly in PNH-AA (0.77763 +/- 0.409153) (p < 0.05). The proportion of Th1 cells in PNH [(16.9136 +/- 6.78899)%], especially in PNH-AA [(22.8000 +/- 5.45244)%], was significantly higher than that in controls [(4.4600 +/- 1.81879)%] (p < 0.05). The proportion of Th2 cells in PNH [(4.7582 +/- 1.98441)%] had no difference from controls [(3.7960 +/- 1.13810)%]. The number of GPI(-) T cells in CD8(+) T cells and CD4(+) T cells were (14.6797 +/- 11.96718)% and (3.9241 +/- 2.46263)% respectively. The expression of CD69 on GPI(+) T cells or GPI(-) T cells in PNH [CD8(+) GPI(+) T cells (17.67881 +/- 8.562493)%, CD8(+) GPI(-) T cells (15.86575 +/- 7.279743)%, CD4(+) GPI(+) T cells (4.65431 +/- 1.984378)%, CD4(+) GPI(-) T cells (4.93181 +/- 1.730001)%]was significantly higher than that in normal controls [CD8(+) GPI(+) T cells (4.68038 +/- 1.216645)%, CD4(+) GPI(-) T cells (1.77339 +/- 0.645259)%] (p < 0.05), but the expression of CD69 on GPI(+) T cells was not different from that on GPI(-) T cells in PNH. It is concluded that high function of cytoimmunity in PNH may be responsible for bone marrow failure but not relates to the existence of PNH clone in T cell population.

Study of the quantity and function of Th17 cells in the blood cytopenic patients with positive BMMNC-Coombs test / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the quantity and function of bone marrow (BM) Th17 cells in the blood cytopenia patients with positive BMMNC-Coombs test (IRP) and to explore the role of Th17 cells in the pathogenesis of the disease.</p><p><b>METHODS</b>Forty-three untreated IRP patients, 34 recovered IRP patients and 13 healthy donors were enrolled in this study. The ratio of IL-23R(+)CD4(+)/CD4(+)cells in BM were examined by flow cytometry(FCM), the levels of IL-6, IL-23, IL-17 by ELISA, and the expressions of RORγt mRNA, STAT3 mRNA in BMMNC by semiquantitive RT-PCR.</p><p><b>RESULTS</b>The ratio of IL-23R(+)CD4(+)/CD4(+)cells [(3.6 ± 3.1)%], the levels of IL-6[(26.21 ± 14.55) µg/L], IL-23[(2.23 ± 0.99) µg/L], IL-17[(2.54 ± 1.33) µg/L] and the expressions of RORγt mRNA (0.25 ± 0.08) and STAT3 mRNA (1.10 ± 0.16) in BMMNC of untreated IRP patients were significantly higher than those of recovered IRP patients[(2.0 ± 1.0)%, (9.08 ± 6.36) µg/L, (0.91 ± 0.76) µg/L, (1.28 ± 0.18) µg/L, 0.12 ± 0.08, 0.97 ± 0.12 respectively] (P < 0.05); there was no significiant difference between those of recovered IRP patients and normal controls [(1.9 ± 1.4)%, (14.63 ± 7.66) µg/L, (1.19 ± 0.98) µg/L, (1.50 ± 0.28) µg/L, 0.07 ± 0.05, 0.95 ± 0.13, respectively] (P > 0.05). In IRP group, there were significantly positive correlations between the ratios of IL-23R(+)CD4(+)/CD4(+)cells and CD5(+)CD19(+)/CD19(+) (P < 0.05), there were significantly positive correlations between the levels of IL-17 and CD5(+)CD19(+)/CD19(+), the quantity of BMMNC-antibody (r = 0.494 and 0.377, respectively) (P < 0.05); and so did between the expressions of RORγt mRNA and the ratio of CD5(+)CD19(+)/CD19(+), the quantity of BMMNC-antibody (r = 0.741 and 0.541, respectively) (P < 0.05), and between the ratio of IL-23R(+)CD4(+)/CD4(+) and the level of IL-17, the expression of STAT3 mRNA (r = 0.438 and 0.448, respectively) (P < 0.05).</p><p><b>CONCLUSIONS</b>There exists increased qunantity and hyperfunction of Th17 cells in the IRP patients which induce B cells hyperfunction and production of autoantibodies against the BM hematopoietic cells. Th17 cells might be a potential new therapeutic target of IRP.</p>

Study on quantity and function of bone marrow macrophages in patients with BMMNC-Coombs Test(+) pancytopenia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the quantity and function of bone marrow (BM) macrophages in patients with BMMNC-Coombs Test(+) pancytopenia (BCTP).</p><p><b>METHODS</b>Sixty-one patients with BCTP, 10 with severe aplastic anemia (SAA) and 13 normal controls were enrolled in this study. The quantity of BM macrophages was measured by FACS and their function was evaluated by phagocytosis test.</p><p><b>RESULTS</b>The number of macrophages, phagocytosis ratio and index of cock's red blood cells (CRBC) in BCTP patients were (0.57 +/- 0.30)%, (37.56 +/- 15.20)%, and 0.75 +/- 0.34, respectively, being significantly higher than those in SAA group \[0.46 +/- 0.08)%, (28.26 +/- 10.46)%, and in 0.59 +/- 0.39\] and in normal control \[0.44 +/- 0.69)% (25.63 +/- 14.75)%, and 0.55 +/- 0.16\] (P < 0.05). The BCTP patients were classified into two subgroups according to the quantity of macrophages: Group A (M(Phi) > or = 0.5%, 34 cases) and Group B (M(Phi) < 0.5%, 27 cases). There were 32 cases (94.12%) with BMMNC-IgG(+) in Group A and only 2 cases (7.41%) in Group B. There were significant differences in phagocytosis ratio and index of macrophages between Group A \[46.62 +/- 13.38)% and 0.91 +/- 0.36\] and Group B \[(28.67 +/- 12.59)% and 0.61 +/- 0.30\] (P < 0.05), while no statistical differences between group B and other two control groups (P > 0.05). Thirty-four BMMNC-IgG(+) patients were further divided into two subgroups: High level (HL) group \[> or = 0.75%, 9 cases (26.47%)\] and Low level (LL) group \[< 0.75%, 25 cases (73.53%)\]. Only one lineage of BMMNC-IgG could be detected in LL group. Among 9 patients in HL group, 8(23.53%) had two lineages of BMMNC-IgG and 1(2.94%) had three lineages. Phagocytosis ratio and index of macrophages were significantly higher in HL group \[(60.22 +/- 12.51)% and 1.23 +/- 0.23\] than in LL group \[(43.32 +/- 9.24)% and 0.84 +/- 0.24\] (P < 0.05). The level of peripheral blood(PB) RBC, HGB and PLT in HL group were significantly lower than in LL group (P < 0.05), while the percentage of Ret, the level of TBIL and the ratio of erythroid of sternal BM in HL group were significantly higher than in LL group (P < 0.05).</p><p><b>CONCLUSION</b>More quantity and stronger function of macrophages are observed in BCTP patients with BMMNC-IgG(+). One of the mechanism of hematocytopenia might be that macrophages activated by IgG autoantibodies phagocytose hematopoietic cells in BM. Macrophages do not involve in damage process of BM in BCTP with IgM or cold autoantibodies.</p>

Variation in complement level and its significance in cytopenia patients with positive BMMNC-Coombs / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the variation of bone marrow complement level in cytopenia patients with positive BMMNC-Coombs test (CBCPC), and probe the role of complement in destroying hematopoietic cells of CBCPC patients.</p><p><b>METHODS</b>One hundred and twenty-four patients with CBCPC and twenty-three healthy donors as controls were enrolled in this study. The levels of CH50, C3, C4, C5b-9 were tested with ELISA. The auto-antibodies on bone marrow hematopoietic cells (BMHC) were examined with flow cytometry.</p><p><b>RESULTS</b>The level of C5b-9 in bone marrow (BM) of untreated CBCPC patients [(119.8+/-54.0) microg/L] was significantly higher than that of recovered patients [(100.7+/-33.4) microg/L] or normal controls [(93.9+/-28.8) microg/L] (P<0.05). The levels of CH50 in BM of untreated or recovered CBCPC patients [(33.3+/-11.5) kU/L, (30.8+/-10.3) kU/L] were significantly higher than that of normal controls [(24.1+/-6.4) kU/L] (P<0.05). The level of C3 in BM of untreated or recovered CBCPC patients [(4.9+/-2.2) mg/L], (5.0+/-3.5) mg/L] was significantly lower than that of normal controls [(7.0+/-5.6) mg/L] (P<0.05). The level of complement in peripheral blood was consistent with that in BM. CH50 in BM of CBCPC patients was negatively correlated with their C3 (r=-0.303, P=.0007) and positively correlated with their C5b-9 (r=0.241, P=0.003) levels. The level of C5b-9 in BM of CBCPC patients was higher in the BMHC-IgM positive group [(117.6+/-55.7) microg/L] than in the BMHC- IgM negative group [(99.2+/-26.2) microg/L] (P<0.05). The positive rate of CD34(+)-IgG or CD34(+)-IgM of CBCPC patients was positively correlated with their C5b-9 level (r=0.593, P=0.000, r=0.326, P=0.049). The reticulocyte percentage (r=0.421, P=0.000) and serum indirect bilirubin level (r=0.230, P=0.032) of CBCPC patients were positively correlated with their CH50 level.</p><p><b>CONCLUSIONS</b>The hematocytopenia of CBCPC patients might be related to the hematopoietic cells destruction caused by auto-antibody activated complements.</p>

Study on DNA-protein Crosslinks in Brain and Testicle Cells Induced by Formaldehyde in Male Mice / 环境与健康杂志

0.05),but as formaldehyde concentration increased,the coefficient of DPC also increased gradually.Higher concentration exposure(1.0,3.0 mg/m~3)resulted in significant elevation of DPC amount compared with the control group(P

Study on the Production of Ergosterol Using Corn Straw Hydrolyzates Fermentation / 中国生物工程杂志

The ergosterols were produced from corn straw hydrolysates fermented by ergosterol yeast,which was obtained from protoplast electrofusion.The effects on the yield of ergosterol were studied in the condition of shaker,such as initial sugar concentration,nitrogen source,pH value and fermentation time.The technical conditions were optimized according to the DPS center-united experimental design principles and the method of response surface analysis with four factors and three levels.The results indicated that the four factors had significant correlation to ergosterol accumulation.The biomass and the ergosterol content could be up to 8.67g/L and 2.37% respectively after cultivated for 32h under optimal technical condition.The structure of ergosterol crystal was characterized by UV,IR and SEM.A new approach of biomass source application was presented.