Objective To analyze the diffusion properties of acute cerebral stroke to understand the effects of infarct and hematoma on diffusion-weighted MR images of patients with acute cerebral stroke. Methods 25 acute cerebral infarcts, 22 intracranial hematomas and 5 hemorrhagic infarctions were studied with CT, conventional and diffusion MR imaging. The DWI and ADC parameters of acute cerebral stroke were analyzed. Results The DWI images of 25 acute cerebral infarcts, 1 hyperacute hematoma, 2 acute hematomas and 3 subacute hematomas were hyperintensity, and that of 8 early subacute hematomas were hypointensity. The DWI images of early stage of 4 chronic hematomas were hyperintensity at center and hypointensity at periphery, and that of late stage were hypointensity. The DWI images of acute hemorrhagic infarctions had both the hyperintensity and hypointensity. Meanwhile, the ADC images of majority of the lesions were hypointensity. Conclusion DWI could reflect the diffusion properties of different types of cerebral strokes and play an important role in evaluating the strokes.

Objective:To evaluate the clinical effect of Chinese medical chiropractic in treating children with cough variant asthma syndrome (CVA) of Lung-Spleen Qi Deficiency.Methods:From October 2017 to October 2018, 60 CVA children with syndrome of lung-spleen qi deficiency were enrolled from pediatric outpatient department of Maoming Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine, and they were randomly divided into the control group and observation group by random number table method, with 30 in each group. The control group was given montelukast sodium granules orally on the basis of conventional treatment, and the observation group was given chiropractic treatment on the basis of the control group. Both groups were treated for 6 weeks and followed up for 2 months. The TCM syndrome scores were performed before and after treatment, and the changes of serum IgA, IgG, IgM and IgE were detected by ELISA. The recurrence was observed and recorded during the follow-up period, and the clinical effect rate was evaluated.Results:The total effect rates of the observation group and the control group were 86.7% (26/30) and 56.7% (17/30), respectively, with significant differences between the two groups ( χ2=6.648, P=0.008). After treatment, the scores of cough, sputum volume, oppression in chest, tongue coating, concomitant symptoms and total scores in the observation group were significantly lower than those in the control group ( t values were 2.639, 2.628, 2.633, 2.627, 2.725, 2.642, respectively, all Ps<0.05). After treatment, the IgA [(2.31 ± 1.20) g/L vs. (1.55 ± 0.72) g/L, t=2.635], IgG [(11.16 ± 3.39) g/L vs. (8.18 ± 3.27) g/L, t=2.751], IgM [(1.72 ± 0.34) g/L vs. (1.27 ± 0.36) g/L, t=2.660] in the observation group were significantly higher than those in the control group ( P<0.05), while the IgE [(51.32 ± 32.62) g/L vs. (74.58 ± 40.77) g/L, t=2.619] in the observation group were significantly lower than those in the control group ( P<0.05). During the follow-up, the recurrence rate was 19.2% (5/26) in the observation group and 70.6% (12/17) in the control group, and the difference between the two groups was statistically significant ( χ2=4.675, P=0.029). Conclusion:Chinese medicine chiropractic can improve the clinical symptoms and serum immune factor levels of children with cough variant asthma syndrome of lung and spleen qi deficiency, reduce the recurrence rate, and improve the clinical efficacy.

Objective To explore the effect of sustained performance on EEG approximate entropy (ApEn).Methods Fifty participants were divided into two groups according to whether they had undergone sustained performance .One day af-ter the sustained-performance group finished executing their task , the resting EEG of nine electrodes ( F3, Fz, F4, C3, Cz, C4, P3, Pz and P4) was acquired in two grups separately , and the ApEn of each electrode was calculated as well . Results The ApEn of F3 and Fz showed an evident decrease as well as randomness of spontaneous brain electrode activity in frontal lobes , especially in the left one .Conclusion The drop of ApEn and the asymmetry of frontal lobes might reflect participants′state of fatigue during sustained performance .

Objective To evaluate the role of three-dimensional multi-tissues (3DMT) CT reconstruction in imaging atlantoaxial and atlantooccipital deformities or traumas.Methods 10 normal volunteers and 42 patients with atlantoaxial and atlanoccipital deformities or traumas underwent spiral CT scan, then three-dimensional (surface shaded display, SSD) CT reconstruction and three-dimensional multi-tissues (3DMT) CT reconstruction.Results SSD and 3DMT reconstruction clearly demonstrated the anatomy of the bony structures and the spatial relationships between the occipitale and atlas, atlas and axis. 3DMT reconstruction respectively demonstrated the occipitale, atlas and axis. 3DMT more neatly demonstrated the deformities, fractures and dislocations of atlantoaxial and atlantooccipital joint than SSD reconstruction. Conclusion 3DMT CT illustrates the anatomical relationship of atlantooccipital and atlantoaxial complex and provides important information of the pathology. 3DMT is prior to SSD reconstruction.

Objective To assess metabolic alterations in the human basal ganglia area during maturation and aging by using 2D chemical shift imaging (2D CSI) of proton magnetic resonance spectroscopy (1H-MRS). Methods Seventy healthy subjects were examined by 2D CSI. 2D CSI imaging acquisition was performed in the bilateral caudate, lentiform and thalamus. 1H-MRS was processed to determine the metabolite ratios, including NAA/Cho, NAA/Cr, Cho/Cr. Seventy healthy subjects were divided into 3 groups:20 to 39 years of age group, 40 to 59 years of age group and 60 to 87 years of age group. The three groups of healthy participants were compared. Results There was a significant decrease with aging in the NAA/Cho ratio in the bilateral lcntiform, thalamus and left caudate, and a significant decrease with aging in NAA/Cr ratio in the bilateral thalamus,right lentiform and left caudate (P<0.05), whereas the Cho/Cr ratio was significantly increased in the bilateral lentiform with aging(P<0.05). Conclusions The results of 1H-MRS show significant changes in the level of metabolites during the process of aging. This technique may play an important role in clinical studies and applications for various conditions of metabolic disorders of the human brain.

OBJECTIVETo construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODSThe total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTSThe libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSIONWe have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; Cell Surface Display Techniques ; Gene Library ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Peptide Library ; Urinary Bladder Neoplasms ; genetics ; immunology

OBJECTIVETo construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).

METHODSThe total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.

RESULTSUsing 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.

CONCLUSIONThe personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.

Amino Acid Sequence ; Child ; Gene Library ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Membrane Proteins ; genetics

OBJECTIVETo screen the HIV-1 entry inhibitors targeting HIV-1 gp120 from the IBS natural product database by virtual screening based on the binding mode of the neutralizing antibody VRC01 with HIV-1 gp120 and investigate the anti-viral activities of the inhibitors and their action mechanisms.

METHODSThe binding interaction of the candidate molecules binding gp120 and changes of the binding free energy were analyzed by MM-PBSA calculation. The anti-HIV-1 activities of the tested compounds were detected by HIV-1 pseudotyped virus, laboratory-adapted HIV-1 and a cell-cell fusion assay. The cytotoxicity of the studied molecules was examined by XTT colorimetric assay. The mechanisms of the anti-viral activities of the candidate molecules were analyzed using enzyme-linked immunosorbent assay.

RESULTSA total of 19 molecules with distinct reduction of the binding free energy after binding with gp120 were screened from 40000 molecules. Among them, NC-2 showed anti-HIV-1 activities against HIV-1 pseudotyped virus and laboratory-adapted HIV-1, and was capable of blocking HIV-1 envelope-mediated cell-cell fusion. The IC50 of NC-2 for inhibiting HIV-1IIIB and pseudotyped HIV-1JRFL infection were 1.95∓0.44 µmol/L and 10.58∓0.13 µmol/L, respectively. The results of ELISA suggested that NC-2 could inhibit the binding of HIV-1 gp120 to CD4 without blocking the formation of gp41 six-helix bundle in vitro.

CONCLUSIONThis computer-based virtual screening method can be used to screen HIV-1 entry inhibitors targeting gp120. Using this virtual screening approach combined with anti-viral activity screening, we obtained a potent HIV-1 entry inhibitor NC-2 with novel structure.

Anti-HIV Agents ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Neutralizing ; pharmacology ; Binding Sites ; Cell Fusion ; Cell Line ; Drug Discovery ; Drug Evaluation, Preclinical ; HIV Antibodies ; pharmacology ; HIV Envelope Protein gp120 ; antagonists & inhibitors ; HIV-1 ; drug effects ; Humans ; Microbial Sensitivity Tests

OBJECTIVE: To explore the inhibitory effect of PSB0739 on the formation of semen-derived amyloid fibrils. METHODS: PAP248-286 (440 μmol/L) was incubated with PSB0739 at different concentrations, and at different time points of incubation, aliquots were taken from each sample for Congo red staining to detect the formation of amyloid fibers. The morphology of amyloid fibrils incubated in the presence or absence of PSB0739 was visualized using transmission electron microscope. The effect of PSB0739 on amyloid fibril formation was determined using virus infection assays at different time points, and the surface charges of amyloid fibril incubated with PSB0739 were calculated using a Zeta potentiometer. The cytotoxicity of PSB0739 in Hela cells was determined using MTT assay. The antiviral effect of PSB0738 against HIV- 1 was evaluated by infection assay. RESULTS: PSB0739 inhibited SEVI fibril formation in a dose-dependent manner . At 48 h of incubation, 220 μmol/L of PSB0739 obviously inhibited the formation of amyloid fibrils in 440 μmol/L of SEVI. Transmission electron microscopy revealed that 220 μmol/L PSB0739 prevented PAP248- 286 (440 μmol/L) from forming amyloid fibrils. PSB0739 antagonized SEVI-mediated enhancement of HIV-1 infection, and 1760 μmol/L of PSB0739 completely reversed the positive charge of SEVI ( < 0.05). PSB0739 below the concentration of 62.5 μmol/L showed no obvious cytotoxicity in Hela cells (>0.05). PSB0739 showed a direct anti-HIV activity with an IC of 21.77±5.15 μmol/L. CONCLUSIONS PSB0739 can inhibit the formation of semen-derived amyloid fibrils .
Amyloid ; chemistry ; drug effects ; Anti-HIV Agents ; pharmacology ; Dose-Response Relationship, Drug ; HIV Infections ; drug therapy ; HIV-1 ; drug effects ; HeLa Cells ; Humans ; In Vitro Techniques ; Microscopy, Electron, Transmission ; Purinergic P2Y Receptor Antagonists ; pharmacology ; Semen ; chemistry

ObjectiveTo investigate the changes of intestinal microbiota in type 2 diabetic diarrhea （T2DD） patients with dampness-heat syndrome and spleen-kidney deficiency syndrome. MethodT2DD patients who were admitted to the Department of Endocrinology Ⅰ of the Affiliated Hospital of Shaanxi University of Chinese Medicine from March 2020 to May 2021 were selected， including 14 patients with type 2 diabetes mellitus （T2DM）， 12 T2DD patients with dampness-heat syndrome， and 13 T2DD patients with spleen-kidney deficiency syndrome. Twelve healthy subjects receiving medical examination were selected as control group. Their body mass index （BMI）， fasting plasma glucose （FPG）， 2 h postprandial blood glucose （2 h PBG） and glycosylated hemoglobin （HbA1c） were compared. Fecal samples were collected for DNA extraction to build a database. High-throughput 16S rDNA sequencing was used to compare the composition of intestinal microbiota and the differential bacteria among the four groups. ResultCompared with the conditions in control group， the levels of FPG， 2 h PBG and HbA1c in the other groups were increased （P<0.05）. Alpha diversity showed no significant difference in species richness， evenness and diversity of intestinal microbiota among the groups. Beta diversity indicated that intestinal microbiota tended to be consistent in each group， and there was no marked difference between groups. The top 5 phylum by relative abundance were Bacteroidetes，Proteobacteria， Firmicutes， Actinobacteria， and Fusobacteria， among which，Bacteroidetes， Proteobacteria and Firmicutes were dominant. Compared with the control group， the three diabetic groups had elevated relative abundance of Bacteroidetes while decreased relative abundance of Firmicutes. The relative abundance of Actinomycetes in spleen-kidney deficiency T2DD group was significantly higher than that in the other groups， and the relative abundance of Firmicutes and Fusobacteria in the dampness-heat T2DD group was significantly lower than that in the other groups. At the genus level， the top 10 bacteria by relative abundance were Phocaeicola， Bacteroides， Pseudescherichia， Prevotella， Bifidobacterium， Faecalibacterium， Fusobacterium， Roseburia， Citrobacter， and Cetobacterium. LEfSe analysis revealed that the relative abundance of Prevotella， Mediterraneibacter， Parabacteroides， and Fusicatenibacter in diabetic patients was remarkably higher than that in healthy patients. Bacteroides and Sutterella might be the characteristic microbiota of T2DD patients with dampness-heat syndrome， while Faecalibacterium， Limosilactobacillus， Eubacterium， Gemmiger， Enterocloster， Alistipes， Parasutterella and Oscillibacter might be the characteristic microbiota of T2DD patients with spleen-kidney deficiency syndrome. ConclusionBacteroides and Parasutterella might be the characteristic microbiota of T2DD patients with dampness-heat syndrome and spleen-kidney deficiency syndrome， respectively. This paper provided reference for studying the mechanism， diagnosis and treatment of modern traditional Chinese medicine for T2DD of dampness-heat type and spleen-kidney deficiency type.