Objective:To explore the effects of Silybin on pulmonary inflammatory injury and macrophage apoptosis in pulmo-nary tuberculosis(TB)rats,and its regulation on death receptor Fas and its ligand FasL.Methods:TB rat model was prepared by tail vein injection of Mycobacterium tuberculosis(Mtb).Rats were randomly separated into model group,Silybin group,Fas overexpres-sion recombinant protein(pcDNA-Fas)group,pcDNA-Fas negative control(pcDNA-NC)group and Silybin+pcDNA-Fas group,with 15 rats in each group,and another 15 rats were selected as normal control group.Acid-fast staining was used to measure infection of lung tissue;HE staining was performed to observe pathological changes of lung tissue;expressions of TNF-α and IL-6 in lung tissue were detected by ELISA;apoptosis rate of alveolar macrophages was detected by flow cytometry combined with Annexin V-FITC/PI staining;expression levels of Fas,FasL,caspase8,caspase3 and macrophage inflammatory protein-2(MIP-2)were detected by Western blot.Results:Compared with normal control group,expressions of inflammatory factors in lung tissue and apoptotic rate of alveolar macrophages were increased in model group,Mtb infection and caseous necrosis in lung tissue were severe,and Fas/FasL-mediated caspase8/3 apoptotic pathway was activated(P<0.05).Compared with model group,expressions of inflammatory factors in lung tissue and apoptosis rate of alveolar macrophages in Silybin group were reduced,Mtb infection and caseous necrosis in lung tissue were alleviated,and the activity of Fas/FasL-mediated caspase8/3 apoptosis pathway decreased(P<0.05).pcDNA-Fas was able to further activate Fas/FasL-mediated caspase8/3 apoptotic pathway,aggravate lung tissue Mtb infection and caseous necrosis,promote inflammatory damage in lung tissue and macrophage apoptosis,and weaken the anti-Fas/FasL activation,anti-inflammatory and anti-apoptotic effects of Silybin(P<0.05).Conclusion:Silybin may play an anti-Mtb infection,anti-apoptosis of lung tissue macro-phages and anti-inflammatory effects by inhibiting the Fas/FasL signaling pathway.

Objective:To investigate the risk factors of diabetic nephropathy (DN) in primary type 2 diabetes mellitus (T2DM) patients and to quantitatively analyze the risk of DN by nomogram modeling.Methods:A total of 1 588 primary T2DM patients from 17 townships and streets in Zhejiang Province were enrolled from June 2018 to August 2018 in this cross-sectional study, with an average age of (56.8±10.1) years (50.06% male) and a mean disease duration of 9 years. The clinical data, biochemical test results, and fundus photographs of all T2DM patients were collected, and logistic regression analysis was used to screen the risk factors of DN. Then, a nomogram model was used to quantitatively analyze the risk of DN.Results:DN occurred in 27.71% (440/1 588 cases) primary type 2 diabetes patients. Hemoglobin A 1c (HbA 1c) ( OR=1.159, 95% CI 1.039-1.292), systolic blood pressure ( OR=1.041, 95% CI 1.031-1.051), serum creatinine (Scr) ( OR=1.011, 95% CI 1.004-1.017), serum globulin (GLOB) ( OR=1.072, 95% CI 1.039-1.105), diabetic retinopathy (DR) ( OR=1.463, 95% CI 1.073-1.996), education level of more than junior high school ( OR=2.018, 95% CI 1.466-2.777), and moderate-intensity exercise ( OR=0.751, 95% CI 0.586-0.961) were influencing factors of DN. Nomogram model analysis showed that the total score of each factor of DN ranged from 64-138 points, and the corresponding risk rate ranged from 0.1-0.9. The nomogram model also predicted a C-index value of 0.753 (95% CI 0.726-0.781) and an area under the receiver operating characteristic curve of DN of 0.753. Internal verification of the C-index reached 0.738. The model displayed medium predictive power and could be applied in clinical practice. Conclusions:HbA 1c, systolic blood pressure, Scr, GLOB, DR, and more than a junior high school education are independent risk factors of DN. Nomogram modeling can more intuitively evaluate the risk of DN in primary T2DM patients.

Objective:To evaluate and improve the performance of the newborn screening program for primary carnitine deficiency (PCD) based on tandem mass spectrometry and to investigate the incidence of PCD and molecular characteristics of SLC22A5 gene in Guangzhou.Methods:A total of 200 180 neonates born in Guangzhou from 2015 to 2019 were enrolled into the newborn screening program for PCD by tandem mass spectrometry at Guangzhou Newborn Screening Center. The positive results of screening for PCD was defined as free carnitine (C0) less than 10 μmol/L with decreased acylcarnitine species in dried blood spots of three to seven days after birth. Screen-positive newborns and their mothers were recalled for another blood spot sample. The diagnosis was confirmed based on decreased levels of C0 and acylcarnitine species in recalled blood spots and genetic analysis in SLC22A5 gene sequencing. The utility of using the sum of propionylcarnitine and palmitoylcarnitine (C3+C16) as a biomarker for acylcarnitine species in newborn screening was retrospectively evaluated. The levels of C0 and (C3+C16) at first screening were compared between newborns with PCD and newborns born to mothers with PCD by independent t test. The variant spectrum and known pathogenic variants carrier rate of SLC22A5 in 2 395 healthy children in Guangzhou Women and Children′s Medical Center through whole exon sequencing were analyzed. Results:Among 200 180 neonates, 239 (0.12%) cases were screen-positive for PCD. A total of 37 patients including 15 newborns and 22 mothers had confirmed PCD. The incidence of PCD was 1/13 345 in newborns and 1/9 099 in mothers, respectively. The positive predictive value of this program was 15.5%. Taking cutoff values of C0<8.5 μmol/L or C0 8.5~9.9 μmol/L with (C3+C16)<2 μmol/L, the number of screen-positive cases would be reduced from 810 to 224 without additional false negative case, when compared with cutoff value C0<10 μmol/L only. Both levels of C0 and (C3+C16) at first screening were not significant difference between newborns with PCD and newborns born to mothers with PCD ((6.2±2.4) vs. (5.0±1.8) μmol/L, (1.4±0.4) vs. (1.2±0.5) μmol/L, t=3.826, 0.326; P=0.058, 0.572). Seven PCD mothers experienced moderate fatigue and dizziness in the morning. One of them presented with cardiomyopathy in pregnancy. Genetic analysis of the SLC22A5 gene showed that p.S467C, p.F17L, p.R254X were the three most common variants in newborns with PCD. In PCD mothers and healthy children, the p.S467C, p.F17L and R399W were the three most common whereas the severe variant p.R254X was rare. The population carrier rate for pathogenic variants was 1 in 65 and the estimated incidence of PCD was about 1/16 500. Conclusions:Newborn screening can detect PCD both in newborns and mothers. Adding a quantitative biomarker (C3+C16) <2 μmol/L into the newborn screening program can improve the PCD screen performance. The severe variant p.R253X was common in PCD newborns but rare in PCD mothers and healthy children, indicating that the current screening program maybe failed to detect all PCD newborns and under-estimated the incidence rate of PCD in Guangzhou.

Objective:To study the inhibitory effect of salidroside on the proliferation of fibroblast-like synoviocytes with rheumatoid arthritis in human(HFLS-RA) induced by tomor necrossi factor-α(TNF-α),and to clarify the molecular mechanism of its control effect on rheumatoid arthritis(RA).Methods:The HFLS-RA were cultured in vitro,then treated with TNF-α and different concentrations of salidroside.The cells were divided into normal control group(0 μg·L-1TNF-α),model control group(10.0 μg·L-1TNF-α)and 12.5,25.0,50.0,and 100.0 μmol·L-1 salidroside groups(10.0 μg·L-1TNF-α+salidroside).The proliferation activity was detected by MTT mehthod;the expression levels of β-catenin,matrix metalloproteinase-7(MMP-7),and Cyclin-D1 in supernatant of the cells were detected by ELISA method;the expression level of β-catenin protein in cells was detected by Western blotting method.Results:Compared with normal control group,the proliferation activity of the HFLS-RA in model control group was significantly increased (P<0.05);compared with model control group,the proliferation activities of the HFLS-RA in 12.5 and 25.0 μmol·L-1 salidroside groups were decreased but there were no significant differences(P>0.05),and the proliferation activities of the HFLS-RA in 50.0 and 100.0 μmol·L-1 salidroside groups were significantly decreased (P<0.05).Compared with normal control group,the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in model control group were increased(P<0.05).Compared with model control group,the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in 12.5 and 25.0 μmol·L-1 salidroside groups were decreased,but there were no significant differences(P>0.05);the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in 50.0 and 100.0 μmol·L-1 salidroside groups were decreased(P<0.05).The Western blotting results showed that the expression level of β-catenin protein in the cell in model control group was higher than that in normal control group(P<0.01);the expression levels of β-catenin protein in the cells in 12.5 and 25.0 μmol·L-1 salidroside groups were lower than that in model control group,but there were no significant differences(P>0.05);the expression levels of β-catenin protein in the cells in 50.0 and 100.0 μmol·L-1 salidroside groups were lower than that in model control group(P<0.05).Conclusion:Salidroside could inhibit the proliferation of HFLS-RA,and its control effect might be related to the regulation of Wnt/β-catenin single pathway.

Objective:To explore the characteristics of immunological reconstitution of T-cell subsets and the role of Cornus,a Chinese medicinal herb on T lymphocytes in mice after irradiation (IR). Methods:Irradiated model mice were exposed to a single dose of X-ray radiation (2. 6 Gy) with or without Cornus treatments. Bloodroutine was examined before or after irradiation. CD3+,CD4+, CD8+ T cells and Th1, Tc1, Th2, Th17, Treg from spleen or peripheral blood were determined by flow cytometry. Results: Total lymphocytes and CD3+T cells including CD4+T and CD8+T cells,were significantly reduced 3 days after irradiation (P<0. 05). The re-constitution of CD3+ T cells ( especially CD8+T cells ) started from 5 days post irradiation, CD4+T cells increased 8 days after irradiation. However,the production of IFN-γ by Th1 or Tc1 cells were evidently decreased compared with control group even 8 days post irradiation (P<0. 05). In contrast to Th1,Th2/Th17/Treg were observably increased in irradiation group (P<0. 05). But the percentage of Th1 was obviously increased, and Th2, Th17, Treg were markedly decreased in cornus treated mice compared with irradiated mice (P<0. 05). Conclusion: The immunological reconstitution of T lymphocytes started from 5 days after total irradiation with rapid recovery of CD8+ T cells but not CD4+T cells. Cornus effectively improved the imbalance of T cell subsets by promoting the proliferation of Th1 and suppressing Th17 and Treg.

Aim To investigate the protective effect of cinnamic aldehyde ( CA ) on hormone-induced osteo-clasts proliferation and bone resorption in vitro and its molecular mechanisms. Methods RAW264. 7 cells induced into osteoclast were treated with RANKL and M-CSF and then were divided into control group, dexa-methasone ( DEX ) group and different doses of CA (11. 6, 23. 2, 46. 4 μg·L-1 ) groups. OCs were ob-served after tartrate resistant acid phosphatase( TRAP) staining. The cell proliferation was determined by MTT assay at different time points. The expression levels of TRACP5 b in cell cultured supernatants were measured by ELISA. RT-PCR technique was applied to examine the transcriptional levels of RANK and NFATc1 . Re-sults In MTT assay, the proliferation of osteoclasts stimulated by dexamethasone was promoted seriously compared with negative control group ( P < 0. 05 ) . Meanwhile, DEX could strengthen the content of TRACP5 b and up-regulate the expressions of RANK and NFATc1 mRNA. After administration of CA, the proliferation was inhibited, while the enhanced expres-sion of TRAP5b was reversed,and the over-expressions of RANK and NFATc1 mRNA were obviously down-regulated in a time-and-dose-dependent manner ( P <0. 05 ) . Conclusion The results suggest that CA in-hibits proliferation and bone resorption of osteoclast in-duced by DEX, which may be mediated by down-regu-lation of RANK and NFATc1 mRNA.

OBJECTIVETo investigate effects of glucose excursion on the oxidative/antioxidative system in subjects with different types of glucose regulation.

METHODSA total of 30 individuals with normal glucose regulation (NGR), 27 subjects with impaired glucose regulation (IGR) and 27 subjects with newly diagnosed type 2 diabetes mellitus (T2DM) were selected and recruited for 3 days' continuous glucose monitor system (CGMS) assessment. The data from CGMS was used to calculate the mean amplitude of glycemic excursion (MAGE), mean blood glucose (MBG) and its standard deviation (SDBG), area under the ROC curve when the blood glucose >5.6 mmol/L within 24 h (AUC 5.6), mean of daily differences (MODD), and mean postprandial glucose excursion (MPPGE). In all groups, the content or activity of malondialdehyde (MDA), total antioxidation capacity (TAOC) and glutathione peroxidase (GSH-Px) were detected.

RESULTSGlucose excursion parameters of subjects with T2DM or IGR were higher than those of NGR subjects (P<0.05 or 0.01). Moreover, Glucose excursion parameters of T2DM subjects were higher than those of IGR subjects (P<0.05 or 0.01). Subjects with T2DM or IGR had significant higher MDA levels and lower GSH-Px/MDA and TAOC/MDA levels compared to NGR subjects (P<0.01). T2DM subjects had even higher MDA levels and lower GSH-Px/MDA levels than IGR (P<0.05 or 0.01). According to the median of normal population for MAGE, T2DM and IGR subjects were divided into MAGE>2.6mmol/L Group and MAGE ≤ 2.6mmol/L Group. MAGE>2.6mmol/L Group had higher levels of MDA and lower levels of GSH-Px/MDA than MAGE ≤ 2.6mmol/L Group (P<0.05). There was no significant difference between the two groups (P>0.05) in terms of the levels of TAOC/MDA. Pearson correlation analysis showed that MDA was positively correlated with FPG, 2hPG, MAGE, and SBP. GSH-Px/MDA was negatively correlated with MAGE and TC. TAOC/MDA was negatively correlated with FPG. Partial correlation analysis showed that the relationship between MDA and MAGE, GSH-Px/MDA, and MAGE remained significant after adjustments for the other differences among groups.

CONCLUSIONGlucose excursion contributed significantly to promoting lipid peroxidation and decreasing antioxidation capacity than chronic sustained hyperglycemia did in the subjects with different types of glucose regulation.

Adult ; Antioxidants ; metabolism ; Blood Glucose ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Oxidation-Reduction