Objective To obtain two types of optical isomers from 1-(Benzamidomethyl)-1,2,3,4-tetrahydro-isoquinoline (BTIQ). Methods ( ±) BTIQ as raw materials , optically pure camphor sulfonic acid as resolution agent was used and repeated resolution in acetone , And the resolution product was detected by specific optical rotation .The product of BTIQ was hydrolyzed , and the ratio of ATIQ was compared with that of the literature .ResuIts After two repeated chemical resolutions , the specific rotations of both enantiomers are no longer changed .It was showed that the products of higher optical purity.The specific rotation of both isomers are -35.65°(CH2Cl2, C=0.5)with the yield of 27.52%,and +35.17°(CH2Cl2, C =0.5) with the yield of 31.55% respectively.The specific rotation value of chiral 1,2,3,4-tetrahydroisoquino line ( ATIQ) which was the hydrolysis product of BTIQ consistent with values reported in the literature .ConcIusion The (-) BTIQ and ( +) BTIQ enantiomers were successfully obtained by the method of resolution , and the yield and optical purity of the obtained products are higher , laying the foundation for the further development of high efficiency , low toxicity of chiral schistosomicide ( praziquantel ) and other containing tetrahydroisoquinoline structure of chiral drugs .

Objective To investigate the immunoregulatory effect of Fasudil-modified macrophages on cell transferred experimental autoimmune encephalomyelitis ( EAE) in a mouse model.Methods Fe-male C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE.The encephalomyelitic mononuclear cells ( MNCs) were isolated from spleen of mice with EAE on day 9 after immunization and treated with or without Fasudil for 72 h in vitro.Several assays including the flow cytometry analysis, Griess reaction and ELISA were performed to analyze the M1 and M2 phenotypes of macrophages, the production of NO and the levels of cytokines, respectively.The cultured MNCs (5×107 cells) were resuspended in 500μl of PBS and transferred into na?ve C57BL/6 recipients via intraperitoneal injection.Two groups including the PBS-MNCs group and the Fasudil-MNCs group were set up.The body weights and clinical scores of the mice in each group were recorded in every other days after the induction of EAE in the recipients.Results The Fasudil treated MNCs affected the induction of EAE in adoptive cell transferred mice.The expression of CD16/32, iNOS and IL-12 on F4/80-macrophages were decreased, while the expression of CD206, CD23 and IL-10 on F4/80-macrophages were increased upon the treatment of Fasudil, indicating that Fasudil im-proved the differentiation of macrophages from M1 to M2 phenotypes.Moreover, Fasudil inhibited the pro-duction of NO and enhanced the expression of Arginase-1.Conclusion Fasudil ameliorated the clinical se-verity of EAE in mice by promoting the transformation of macrophages from M1 to M2 phenotype.

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor WAR 5 on the experimental autoimmune encephalomyelitis (EAE) and its possible mechanism.METHODS: Female C57BL/6 mice were randomly divided into EAE group and WAR5 group.EAE model was induced by the application of MOG 35-55 peptide.WAR5 was in-jected intraperitoneally every other day from post-immunization (PI) day 3 to PI day 27.The clinical score and body weight were recorded every other day .On PI day 28, the animals were sacrificed and spinal cords were obtained for HE and mye-lin staining .The splenocytes were isolated and the expression of CD 16/32 and CD206 were analyzed by flow cytometry . The protein extracts from the brains and spinal cords were collected for the measurement of inducible nitric oxide synthase ( iNOS) by Western blotting .RESULTS:The administration of WAR 5 delayed the onset of EAE and attenuated the clini-cal symptoms .The results of the pathological examination revealed that WAR 5 inhibited the infiltration of inflammatory cells and improved myelination in spinal cords , accompanied with the poralization of M 1 macrophages to M2 phenotype in the spleen.WAR5 inhibited the expression of iNOS in the central nervous system , especially in the spinal cords .CON-CLUSION:The therapeutic effect of WAR5 on EAE may be related to the shift of M1 macrophages to M2 phenotype and inhibition of inflammation in the central nerve system .

AIM:To explore the therapeutic effect of Buyang-Huanwu decoction (BYHWD) on experimental au-toimmune encephalomyelitis ( EAE) and its immunoregulatory effect on monocyte-macrophages .METHODS: Chronic EAE was induced by myelin oligodendrocyte glycoprotein peptide fragment 35-55 ( MOG35-55 ) in the female C57BL/6 mice, which were randomly divided into saline group and BYHWD group .On day 3 after immunization , the mice in BYHWD group were orally administrated with BYHWD , while normal saline was given to the control mice .The clinical score and body mass were recorded every other day .At day 17 after immunization , the mice were sacrificed and spinal cords were obtained for HE staining and myelin staining .The M1 and M2 macrophage phenotypes of splenic cells were detected by flow cytometry and immunofluorescence staining .The protein expression of iNOS , TNF-α, arginase and IL-10 in the spinal cord macro-phages was determined by Western blotting .RESULTS:BYHWD delayed the onset of EAE , reduced the clinical scores of EAE, inhibited the inflammatory cell infiltration and demyelination in the spinal cord , and promoted the conversion of M 1 macrophages into M2 phenotype in the spinal cord and spleen .CONCLUSION:BYHWD intervention attenuates the be-havioral and pathological changes in the EAE mice , and its mechanism may be related to the macrophage conversion .

Objective:To investigate the relationship between procalcitonin(PCT) and insulin resistance in diabetic foot infection.Methods:Sixty patients with diabetic foot infection hospitalized in the Fifth People's Hospital of Datong from March 2015 to March 2017 were selected and divided into three groups according to the value of PCT: slightly elevated group(L group, n=18), moderately elevated group(M group, n=21), highly elevated group(H group, n=21). Another 20 patients with type 2 diabetes mellitus were collected as control group.The PCT, C-reactive protein(CRP), white blood cell count(WBC), fasting and postprandial blood glucose, fasting insulin and insulin resistance index(HOMA-IR) were calculated and compared. Results:In the control group, the levels of PCT, HOMA-IR, CRP and WBC were (0.14±0.12)μg/L, (17.70±8.86), (32.90±24.19)mg/L, (8.01±2.21)×10 9/L, respectively, which in the L group were (0.31±0.14)μg/L, (20.42±9.71), (50.85±27.81)mg/L, (9.95±3.35)×10 9/L, respectively, which in the M group were (1.11±0.52)μg/L, (24.08±14.09), (64.31±40.21)mg/L, (10.86±2.25)×10 9/L, respectively, which in the H group were (5.31±3.04)μg/L, (31.73±14.13), (72.29±50.26)mg/L, (12.51±5.51)×10 9/L, respectively, and there were statistically significant differences among the four groups( F=50.744, 5.195, 4.303, 5.252, all P<0.01). With the increase of PCT, the levels of WBC, CRP and HOMA-IR were increased, and HOMA-IR was positively correlated with PCT( r=0.265, P=0.017). Conclusion:Serum level of PCT has correlation with infection degree of diabetic foot and is positively correlated with insulin resistance.

Objective:To compare serum procalcitonin and fibrinogen degradation product levels between type 2 diabetes mellitus patients with Escherichia coli bloodstream and urinary tract infections. Methods:The clinical data of 82 type 2 diabetes mellitus patients with Escherichia coli infections who received treatment between December 2014 and December 2019 in the First Affiliated Hospital of Datong University (The Fifth People's Hospital of Datong) were retrospectively analyzed. These patients were assigned to bloodstream infection ( n = 40) and urinary tract infection ( n = 42) according to the way of Escherichia coli infection. Serum procalcitonin and fibrinogen degradation product levels, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, C-reactive protein, white blood cell count, D-Dimer level, antithrombin III activity, and electrolytes were determined and compared between the two groups. Correlation between procalcitonin and other variables was analyzed. Multiple linear regression analysis was performed with procalcitonin level as a dependent variable and other relevant indexes as independent variables. Results:Body temperature, white blood cell count, neutrophil count, monocyte count, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, procalcitonin level, C-reactive protein level, fibrinogen degradation product level, and D-Dimer level in the bloodstream injection group were (39.49 ± 0.64) ℃, (14.92 ± 11.78) × 10 9/L, (13.39 ± 11.60) × 10 9/L, (0.72 ± 0.36) ×10 9/L, (14.86 ± 10.52), (199.15 ± 160.69), (22.81 ± 17.86) μg/L, (133.44 ± 63.63) mg/L, (49.71 ± 41.44) mg/L, (16.56 ± 12.20) mg/L, respectively, which were significantly higher than those in the urinary tract infection group [(37.12 ± 1.20) ℃, (9.04 ± 3.95) × 10 9/L, (6.25 ± 4.02) × 10 9/L, (0.42 ± 0.29) × 10 9/L, (3.67 ± 3.34), (120.01 ± 44.08), (4.46 ± 8.69) μg/L, (39.22 ± 22.16) mg/L, (3.81 ± 3.41) mg/L, (0.84 ± 0.75) mg/L), t = 7.356, 2.578, 3.162, 2.958, 5.538, 2.591, 2.810, 4.825, 2.902, 2.375, all P < 0.05]. Platelet count, lymphocyte count, blood sodium level and antithrombin Ⅲ activity in the bloodstream infection group were (167.50 ± 104.93) × 10 9/L, (1.06 ± 0.58) × 10 9/L, (130.89 ± 6.50) mmol/L, (57.88 ± 16.28)% , which were significantly lower than those in the urinary tract infection group [(239.40 ± 82.52)× 10 9/L, (2.14 ± 0.71) × 10 9/L, (138.46 ± 5.96) mmol/L, (90.11 ± 8.90)%, t = -2.853, -6.313, -4.046, -7.350, all P < 0.05]. Correlation analysis revealed that serum procalcitonin level was positively correlated with body temperature ( r = 0.387), white blood cell count ( r = 0.355), neutrophil count ( r = 0.368), C-reactive protein ( r = 0.605), fibrinogen degradation product level ( r = 0.616), D-Dimer level ( r = 0.486) (all P < 0.05), and it was negatively correlated with sodium level ( r = -0.319) and antithrombin Ⅲ activity ( r = -0.465) (both P < 0.05). Multiple linear regression analysis results revealed that fibrinogen degradation product level and body temperature were greatly correlated with procalcitonin level. Conclusion:Inflammatory indicators procalcitonin level, body temperature, white blood cell count, neutrophil count, C-reactive protein, fibrinogen degradation product level and D-Dimer level were remarkably higher in type 2 diabetes mellitus patients with Escherichia coli bloodstream infection than those in type 2 diabetes mellitus patients with Escherichia coli urinary tract infection. Procalcitonin level was greatly correlated with body temperature and fibrinogen degradation product level.

OBJECTIVETo identify a novel hemoglobinopathy applied by direct sequencing and clone sequencing.

METHODSEDTA anticoagulated blood of proband and his parents were analyzed by hematology analyzers and Capillarys hemoglobin electrophoresis (CE). Then thalassemia genetypes were screened by gap-PCR and reverse dot blot (RDB). Proband was suspected with abnormal hemoglobin combine alpha beta compound thalassemia. The mutation of beta-globin was identified by direct sequencing and clone sequencing.

RESULTSHb analysis showed that probands Hb A2 variant was eluted in Z (C) zone and his father's in Z (A2) zone on CE,and proband's mother elevated HbA2 of 4.6%. Screened by RDB, the proband was CD71-72(+A) homozygote and showed the mismatch with his parents. Through direct sequencing and clone sequencing, we deduced that our proband inherited the mutations of HBB c.[219T>A;220G>T] from his father and inherited the Southeast-Asian deletion and HBB c.216-217insA from his mother.

CONCLUSIONA novel double heterozygote of HBB c.[219T>A; 220G>T] was identified in south China. This mutation enriches the beta-thalassemia gene mutation spectrum in Chinese population.

Asian Continental Ancestry Group ; genetics ; Child, Preschool ; Hemoglobins ; genetics ; Hemoglobins, Abnormal ; genetics ; Heterozygote ; Humans ; Male ; Mutation ; genetics ; Pedigree ; Thalassemia ; genetics ; beta-Globins ; genetics

Objective To investigate whether astragaloside ( AST) Ⅳ could inhibit lipopolysac-charide (LPS)-induced activation of astrocytes and the possible mechanism. Methods Effects of different concentrations of AST Ⅳ on astrocyte viability were determined by MTT to select the optimum concentration for the following experiments. Primary astrocytes were induced by LPS to construct the inflammatory model. Astrocytes were divided into three groups: PBS, LPS and LPS+ASTⅣ(LPS+ASTⅣ) groups. The release of nitric oxide (NO) was detected by Griess method. Expression of glial fibrillary acidic protein ( GFAP), an astrocyte marker, and TLR4 were analyzed by immunohistochemistry and Western blot. Cytokines of IL-6, TNF-α, IL-4 and IL-10 in the supernatants of cell culture for 24 h collected after stimulation were meas-ured by ELISA. Results ASTⅣsignificantly inhibited the LPS-induced activation of astrocytes and NO re-lease (P<0. 01), suppressed the expression of TLR4 (P<0. 05) and reduced the secretion of IL-6 (P<0. 01) and TNF-α (P<0. 01), but increased the secretion of IL-4 and IL-10 (P<0. 05 and P<0. 01). Con-clusion AST Ⅳ could significantly inhibit the LPS-induced activation of astrocytes and suppress inflamma-tory responses, and the possible mechanism might be related to reduced secretion of inflammatory factors af-ter blocking the expression of TLR4.

Objective:To investigate the efficacy of the Mini-Mental State Scale (MMSE) versus the Montreal Cognitive Assessment Scale (MoCA) in screening cognitive impairment in patients with a lacunar cerebral infarction. Methods:138 eligible patients who received treatment in the Affiliated Hospital of Shanxi Datong University from January 2018 to October 2019 were recruited for this study. They received cognitive function evaluation by the MMSE and MoCA. These patients were grouped according to the median number of age or the median number of years of education. The sensitivity and consistency of the MMSE versus MoCA in screening cognitive impairment in patients with a lacunar cerebral infarction were analyzed using the χ2 test. The total cognitive scores of the MMSE and MoCA, and the scores of each cognitive domain such as memory, execution, visual space, attention, language, and orientation, were compared between groups using multiple linear regression analysis. Results:The sensitivity of MoCA in screening for cognitive impairment in low-age, high-age, low-year-education, and high-year-education groups and the whole population of patients with a lacunar cerebral infarction was 76.5%, 75.7%, 74.2%, 77.8%, 76.1%, respectively, which were significantly higher than those of MMSE (44.1%, 65.7%, 60.6%, 50.0%, 55.1%, χ2 = 12.17, 13.13, 9.33, 15.75, 23.86, all P < 0.01). The Kappa coefficients of low-age, high-age, low-year-education and high-year-education groups were 0.336, 0.391, 0.358, 0.389, and 0.373, respectively, all of which were less than 0.4 (all P < 0.01). These findings suggest that the consistency of the two scales in screening cognitive impairment is poor. The cognitive impairment detection rate by the MMSE was significantly higher in the high-age group than in the low-age group (65.7% vs. 44.1%, χ2 = 6.50, P < 0.05). The total cognitive scores of MMSE and MoCA and the scores of memory, execution, visual space, attention, language, and orientation in patients with a lacunar cerebral infarction were significantly lower in the high-age group or low-year-education group than in the low-age group ( tMMSE = 3.61, 2.49, 3.12, 4.26, 1.70, 3.69, 2.24, all P < 0.01; tMoCA = 3.83, 1.75, 3.28, 3.80, 2.21, 4.08, 2.52, all P < 0.05) or high-year-education group ( tMMSE = -2.87, -2.32, -0.85, -2.54, -0.73, -2.57, -2.96, all P < 0.01; tMoCA = -2.95, -1.12, -3.39, -1.54, -1.52, -3.09, -3.02, all P < 0.05). Conclusion:Combined application of MMSE and MoCA has a high clinical value in screening cognitive impairment in patients with a lacunar cerebral infarction. High-age patients with a lacunar cerebral infarction who receive low-year education have memory, execution, visual space, attention, language, and orientation impairments.

Objective:To explore mechanism of Fasudil reducing Aβ1-42 induced BV2 cell injury based on NLRP3 inflamma-some.Methods:BV2 cells were divided into:normal control group,Aβ stimulation group,Aβ+Fasudil intervention group,Aβ+MCC950(NLRP3 inhibitor)intervention group.Cell morphology was observed under microscope.Cell activity was determined of by CCK8.NO release was measured by Griess.NLRP3,caspase 1 and IL-18 expressions were detected by immunofluorescence staining.NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were detected by Western blot.Results:Compared with normal control group,BV2 cells in Aβ stimulation group were activated and showed amoeba-like shape,cell activity was decreased,NO production was increased,NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were increased.Fasudil intervention and MCC950 intervention inhibited cell injury induced by Aβ1-42 in which BV2 cell morphology tended to be normal,cell activity was increased,while produc-tion of NO was reduced,and NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were down-regulated,there was no significant difference between Fasudil intervention group and MCC950 intervention group.Conclusion:Fasudil may alleviate Aβ1-42 induced BV2 cell injury and inflammatory reaction by inhibiting NLRP3 inflammasome activation.