In order to solve the difficucties of renaturation and immunogenicity of new bifunctional fusion protein GAD,inclusion bodies of GAD were modified by PEG-maleimide.Conformational changes of the modified GAD were compared by circular dichroism and tryptophan fluorescence spectroscopy.The biological activity was verified by oral glucose tolerance test and lipid scavenging.The results showed that PEG-maleimide completed the specific-point modification of GAD,and improved its refolding efficiency.The secondary and tertiary structures of mPEGylated GAD were consistent with that of GAD.PEG-GAD has significant hypoglycemic and lipid-lowering effects (P ＜0.001) with longer half life in vivo and lower immunogenicity (P ＜0.01).This study provides effective strategies for the development of strongly hydrophobic peptide drugs.

Based on ninety three acetylcholinesterase inhibitors (AChEIs) which have the same mechanism of action but are different in structural characteristics, the pharmacophore model for acetylcholinesterase inhibitor was constructed by the CATALYST system. The optimal pharmacophore model with three hydrophobic units, a ring aromatic unit and a hydrogen-bond acceptor unit were confirmed (Weight=3.29, RMS=0.53, total cost-null cost=62.75, Correl=0.93, Config=19.05). This pharmacophore model will act on the double active site of acetylcholinesterase and is able to predict the activity of known acetylcholinesterase inhibitors that are used for clinical treatment of Alzheimer's disease (AD), and can be further used to identify structurally diverse compounds that have higher activity treating with Alzheimer's disease (AD) by virtual screening.

Objective:To explore the cytogenetic characteristics of hypospadias in children by karyotype analysis.Methods:From June 2008 to May 2018, 45 children with hypospadias in Tianjin Children's Hospital had cytogenetic abnormalities. Their median age was 10 months(range 3 hours to 5 years old). Of the 45 cases, 20 were proximal hypospadias, 1 was middle hypospadias. All 24 cases had varying degrees of genitourinary malformations. Among them, 15 cases had unilateral or bilateral cryptorchidism, 5 cases had scrotal division, 3 cases had penile scrotal transposition, 3 cases had small penis, 3 cases had indirect inguinal hernia, 1 case had repeated urethra, 1 case had hydrocele and 1 case had concealed penis. To the other systemic malformations, there was 1 with cleft lip and palate and 1 with congenital heart disease. G-banding karyotype analysis of peripheral blood lymphocytes was performed in all 45 cases.Results:Among the 45 cases of hypospadias with abnormal karyotypes, with an abnormal rate of 14.0%, 28 cases (62.22%) had sex chromosome abnormalities, including (47, XXY), (46, XX/47, XXY), (45, X0/47, XYY), etc. Sexual inversion occurred in 8 cases (17.78%), all of which were 46, XX. There were 4 autosomal abnormalities (8.89%), including (46, XY, 9p+ ), (46, XY, 10p+ ) and (46, XY, 1q+ ). Chromosome polymorphism was found in 4 cases (8.89%), including [46, XY, inv(9)] and [46, XY, 16qh+ ], and the equilibrium translocation of 1 case (2.22%) was [45, XY, -21, -22, + t(21; 22)]. Among the 45 cases, 8 sex reversal children with (46, XX) chromosome karyotype were all proximal hypospadias.Conclusions:Children with hypospadias may be associated with chromosomal karyotype abnormalities, including sex chromosomal abnormalities, autosomal abnormalities, chromosome polymorphism and balanced translocation. Among them, sex chromosome abnormality was the most common and balanced translocation was the least.

OBJECTIVE To explore the origin and morphological features of small supernumerary marker chromosomes (sSMCs) in Turner syndrome. METHODS For 5 cases of Turner syndrome with a sSMC identified by conventional G-banding, dual-color fluorescence in situ hybridization (FISH) was applied to explore their origin and morphological features. RESULTS Among the 5 cases, 3 have derived from the X chromosome, which included 2 ring chromosomes and 1 centric minute. For the 2 sSMCs derived from the Y chromosome, 1 was ring or isodicentric chromosome, while the other was an isodicentric chromosome. CONCLUSION The sSMCs found in Turner syndrome have almost all derived from sex chromosomes. The majority of sSMCs derived from the X chromosome will form ring chromosomes, while a minority will form centric minute. While most sSMC derived from Y chromosome may exist as isodicentric chromosomes, and a small number may exist as rings. For Turner syndrome patients with sSMCs, dual-color FISH may be used to delineate their origins to facilitate genetic counseling and selection of clinical regime.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Environmental Monitoring ; methods ; Toluene ; analogs & derivatives ; analysis ; Workplace

Objective To study the relationships between the standing balance and walking ability of hemiplegic stroke survivors.Methods Eighty-eight post-stroke hemiplegic patients who could walk independently for more than 10 metres were selected into a patient group,while 88 healthy counterparts were recruited into a control group.Descriptors of the gait and balance function of both groups were collected using a gait and balance training and evahuation apparatus (Model:AL-600).The gait parameters were step width,walking speed,step length asymmetry (SLA),swing time asymmetry (SWTA) and stand time asymmetry (STA).The balance parameters studied were total trajectory length of the center of plantar pressure (COPD),the average left and right deviation of the center of plantar pressure (COPD-X) and the average anterio-posterior deviation of the center of plantar pressure (COPD-Y).Differences in indexes of gait and balance function between the two groups were analyzed using t-tests,and the relationships between the gait and balance indicators were analyzed using Pearson correlation coefficients.Results The walking speed,step width,COPD,COPD-X,COPD-Y,SLA,SWTA and STA of the patient group [(37.64± 18.29)cm/s,(14.45±4.17)cm,(66.75±29.04)cm,(2.04±1.41)cm,(2.48±1.28)cm,(1.30±0.46),(1.65±0.67) and (1.18±0.16),respectively] all increased significantly more compared to the control group [(90.76±14.72)cm/s,(8.70±2.62) cm,(27.84±6.54) cm,(1.30±0.53) cm,(1.68±0.40) cm,(1.07±0.06),(1.07±0.08) and (1.05±0.06),respectively],though the walking speed was significantly slower than that in the control group (P＜0.05).The patient group's average COPD-X showed slightly and moderately negative correlation with their walking speed,step width,SLA and SWTA (P＜0.05).COPD-Y was weakly related with step width (P＜0.05).Conclusion The standing balance of hemiplegic patients after stroke is related to their walking speed,step width and gait asymmetry.Especially significant correlation is observed between standing balance parameters such as COPD-X and gait parameters.

Objective To investigate the role of CD4 ~+ CD25~+ regulatory T lymphocytes (Treg)in modulating the cellular immune response and pathogenesis of murine pulmonary tuberculosis.Methods Inactivation of Treg was achieved by intraperitoneal injection anti-CD25 (clone PC61,50 μ/mouse) in PC61 group, and rat-IgG (50 μ/mouse) was injected intraperitoneally in control group. All the mice were inoculated intravenously with H37Rv 0. 1 mL (1 × 10~6 CFU) 3 days after Treg inactivation. The effects of Treg inactivation in different tissues were analyzed by flow cytometry. The cellular immune response, pulmonary histopathology and bacterial load were determined in vitro at different time points. The data were compared using homogeneity of variance F test and non-paired t test. Results In spleen, the percentages of Treg/CD4 T lymphocytes in PC61 group and control group were (21. 13± 3. 58)% and (30. 42± 4. 20)%, respectively at day 10 of inoculation (t = 2. 38, P < 0. 05), and those were (16. 12 ± 1. 26)% and ( 17. 34± 1. 62)%,respectively at day 30 of inoculation (t = 0. 84,P>0. 05). The percentages of Foxp3~+/CD4~+ T lymphocytes in PC61 group and control group were (32. 07 ± 3. 95)% and (60. 55 ± 5. 48)%,respectively at day 10 of inoculation (t = 5. 96, P<0. 05). Similar results were achieved in the peripheral blood. Bacillus calmette-guerin (BCG)-specific 1L-17 (ng/L) secreted by murine spleen cells in PC61 group and control group at day 10, 30 and 60 of inoculation were 5. 1± 0.9 vs 0, 43. 1± 10.0 vs5. 9± 2. 8 and 124.8 ± 5.8 vs 102. 5±8. 1, respectively (t = 7. 90, t=5. 10,t = 3. 19; all P<0.05); those of BCG-specific IFN-γ (ng/L) were 28. 4 ± 8. 2 vs 4. 0±1. 3, 685. 9± 128. 6 vs418. 7±20.4 and 310.9±119. 7 vs 32. 8±7. 5, respectively(tO = 4. 21,t = 8. 43, t = 3. 27; all P<0.05);those of TNF-a (ng/L) were 38. 6±5.0 vs 16. 3±4. 0, 112. 9 ±12. 3 vs 71. 5±12. 6 and 86. 2±8. 2vs0, respectively(t = 4. 95, t=3. 33,t/=14.8; all P<0. 05). The lung bacterial load at day 10 of inoculation in PC61 group was lower than that in control group (t = 4. 63, P < 0. 01), but the differences were not significant thereafter. The changes of lung histopathology at late stage of infection (day 120) in PC61 group were less severe than those in control group. Conclusions Murine Tregs increase dramatically after Mycobacterium tuberculosis infection. Treg could inhibit the specific cellular immunity against Mycobacterium tuberculosis, and therefore, may facilitate the persistent infection of Mycobacterium tuberculosis and development of tuberculosis.

Four impurities were isolated from raw material of clindamycin phosphate (CP), and their structures have been determined. LC-MS was used to determine the molecular weights of the impurities in the raw material of CP. Reversed-phase preparative HPLC was used to prepare them, and their chemical structures were identified by HR-MS and NMR. The four unknown impurities were determined as clindamycin-B-phosphate (1), clindamycin-2,4-diphosphate (2), 3',6'-dehydro clindamycin phosphate (3), epi-clindamycin phosphate (4). Impurity 1 has been included in BP and EP, while 2, 3 and 4 have not. The impurities 2, 3, 4 are first separated from raw material of CP.

Objective To investigate plasma level of soluble intercellular adhesion molecule-1 (sICAM-1)and soluble E-slectin (sES) in patients with Keshan disease (KD) accompanied by hypertension,and to further their relation to systolic blood pressure (SBP),diastolic blood pressure (DBP) and ejection fraction (EF) of left ventricle.Methods Patients with Keshan disease investigated in 2007 in Fuyu County,Heilongjiang Province were divided into two groups according to whether they were accompanied by hypertension or not:KDP group (n =17),the patients were diagnosed as Keshan disease accompanied by hypertension;KDN group (n =14),the patients were only diagnosed as Keshan disease.Healthy volunteers lived in the same area as the patients were included in the control group (CON group,n =10).The clinical data (such as age,sex and past medical history),SBP and DBP were recorded.EF was detected by echocardiography.Plasma sICAM-1 and sES concentrations were measured by ABC-enzyme-linked immunosorbent assay technique.Results Plasma level of sICAM-1 in the subjects was significantly elevated in KDP group as compared with that in CON group and KDN group [(399.84 ± 99.06) vs (323.06 ± 61.27) μg/L,P ＜0.05;(399.84 ± 99.06) vs (268.27 ± 72.88) μg/L,P ＜ 0.01].However,there was no statistical significance in plasma sICAM-1 between the KDN group and control group (P ＞ 0.05).The difference of plasma sES between the control group,KDN group and KDP group did not reach statistical significance [(76.31 ± 23.17),(72.26 ± 20.15) and (90.21 ± 19.21) μg/L,F =3.236,P ＞ 0.05].As compared with the KDN group,SBP and DBP were obviously higher in KDP group [(169 ± 27) and (121 ± 10) mmHg,(102 ± 17) and (81 ± 6) mmHg,1 mmHg =0.133 kPa,F =6.376,4.300,all P ＜ 0.01],and EF was not significantly increased [(58 ± 14)％ and (55 ± 14)％,F =0.789,P ＞ 0.05].Furthermore,correlation analysis showed plasma sICAM-1 was positively correlated with SBP (r =0.540 6,P ＜ 0.01)and DBP (r =0.461 2,P ＜ 0.01),but not with EF of left ventricle in subjects whir Keshan disease (r =0.073 4,P ＞0.05).Conclusion These data shows slCAM-1 may play a role in the development of Keshan disease accompanied by hypertension,providing an potential index for its risk assessment.

AIM: Research on the change of the saponins constituents of Radix Bupleuri after being processed has been carried out. METHODS: Saikosaponin a、c、d and b2 are used as the marker ingredients; the change of saponins constituents,both after procession and saponated action,have been determined by HPLC. RESULTS: After being processed,the content of saikosaponin b2 has a significant increase,saikosaponin a,saikosaponin c,saikosaponin d and saikosaponin a + c + d all have slight decrease. After the saponated action,the content of saikosaponin a + c + d has a little change,and saikosaponin b2 has increased significantly. CONCLUSION: The change rules of saponin compounds in processed Radix Bupleuri have been revealed.