italic>α-Conotoxin ArIB[V11L,V16D] is currently the most optimal selective inhibitor of α7 nicotinic acetylcholine receptor (nAChR) known. In order to explore chemical modification methods and enrich its application in targeting nAChR, this study utilized the linker to covalently connect camptothecin and 7-amino-4-methylcoumarin to the [2,4] disulfide bond of ArIB[V11L,V16D]. Therefore, two peptide-drug conjugates (PDCs), ArIB[V11L,V16D]-5 and ArIB[V11L,V16D]-6, and one fluorescent-labeled peptide, ArIB[V11L,V16D]-7 were constructed. Cytotoxicity evaluation showed that the IC₅₀ values against non-small cell lung cancer cell line A549 of the two PDCs were respectively 1.3 and 4.1 times of camptothecin, indicating slight reduction in activity at the cellular level which was related to the linker structure. Fluorescence spectrum scanning revealed that the excitation and emission wavelength of the fluorescent-labeled peptide were 340 nm and 403 nm respectively, and the fluorescence features of 7-amino-4-methylcoumarin as a marker were retained without fluorescence quenching. This modification strategy laid a solid foundation for the further application of α-conotoxin ArIB[V11L,V16D] in PDCs and fluorescent probes.

Objective To analyze and determine the clinical, molecular pathology and genetic features of a Chinese family with dystrophinopathy. Methods Clinical data of the proband and his family members were collected. Immunohistochemistry staining was performed on muscular biopsy tissues with antimerosin, emerin and the N, C and central rod domains of dystrophin. Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes. Multiplex ligation-dependent probe amplification (MLPA) was used to test Duchenne muscular dystrophy (DMD) gene to determine the ways and sites of genetic mutation, and analyze the relationships between genotype and phenotype. Results Patients from this family were clinically diagnosed as muscular dystrophy, and they presented serious manifestations although the immunohistochemistry analysis for the proband exhibited partial loss of dystrophin staining, and positive expression with merosin and emerin. Further test with MLPA detected the loss of exons 45--54 in DMD gene in the proband, while his mother had heterozygositic loss in exons 45--54. Conclusions The losses of exons 45--54 in the proband are all derived from his mother, who carries genetic mutation with normal phenotype. He has been diagnosed as dystrophinopathy. At the same time, his partial loss of dystrophin is not parallel to the out-of-frame mutation of the gene and his severe clinical manifestations. Abnormal expression of dystrophin is the pathological basis for dystrophinopathy phenotype. Its clinical outcome depends not only on the degree of the protein expression, but also on the function of the sites where the DMD gene less occurs.

Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

Altitude ; Altitude Sickness ; Animals ; Hypoxia ; Iron ; Mice ; Spleen

Objective To study the alterations of serum amino acid level in patients with short bowel syndrome (SBS). Methods Blood levels of 17 amino acids were measured in 17 SBS patients on admission to the hospital, and 15 healthy volunteers. Results Compared with the control group, serum concentrations of Val, Leu, Ile, Lys, Met, Ser, His, Cys in patients with SBS significantly decreased ( P

Objective To study the effects of the rehabilitation robot-assisted task-oriented training on the hand function in patients after stroke. Methods From June, 2015 to September, 2016, 35 inpatients suffering from stroke were randomly allocated to control group (n=17) and trial group (n=18). Based on the routine rehabilitation, the trial group accepted robot-assisted task-oriented training, while the control group accepted therapist-assisted task-oriented training, for two weeks. They were measured the active range of motion (AROM) of fingers, assessed with fingers motor of Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) invovled with hands before and after train-ing. Results The inpatients dropped three in the control group, two in the trial group. AROM of extension and flexion of all the fingers, the AROM of extension and total of three fingers of thumb, index and middle, and the total AROM of each finger improved in the trial group af-ter training (t>2.937, P<0.05), while the AROM of extension and flexion of all the fingers, AROM of extension, flexion and total of the fin-gers of thumb, index and middle, total AROM of the fingers of thumb, index and little improved in the control group after training (t>2.528, P<0.05);the AROM of extension and total of the fingers of thumb, index and middle, and the total AROM of fingers of thumb and index im-proved more in the trial group than in the control group (t>2.535, P<0.05). The scores of mass flexion, mass extension, opposition, cylinder grip, spherical grip and total score of FMA improved in the trial group after training (Z>2.000, P<0.05), while the scores of mass extension, opposition and the total score of FMA improved in the control group after training (Z>2.000, P<0.05). There was no significant difference between the two groups on the items and total scores after training (P>0.05). The scores of feeding, dressing, toilet transfers, bathing, groom-ing of MBI and the total score of them improved in the trial group after training (Z>2.041, P<0.05), while the total score of MBI improved in the control group after training (Z=-2.527, P<0.05). There was no significant difference between the two groups in the items and total scores after training (P>0.05). Conclusion The rehabilitation robot-assisted task-oriented training can improve AROM of hemiplegic fingers and grip function.

Objective To study the effect of intensive trunk muscle training on balance and walking in pa- tients with hemiplegia caused by stroke.Methods A total of 90 stroke patients were recruited and randomly divid- ed into a treatment group(45 cases)and a control group(45 cases).All the patients were given conventional reha- bilitation training.Meanwhile,intensive trunk muscle training was also administered for those in the treatment group as well.The trunk control function,balance ability and walking ability were assessed by using the trunk control test, Berg Balance Scale and the balance subscale of the Fugl-Meyer physical performance test,and Holden's functional ambulation classification,respectively,before and after 6 weeks of training.Results It was found that all the pa- tients scored better with the trunk control test,Berg's balance scale,the balance subscale of the Fugl-Meyer physical performance test and Holden's ambulation classification after treatment,and there were significant differences between the two groups after treatment(P

OBJECTIVETo investigate the effects of electro-acupuncture (EA) on the expression of triggering receptor expressed on myeloid cell (TREM)l in ankle joint synovial tissue of acute gouty arthritis (AGA) rats.

METHODSForty male SD rats were randomly divided into 4 groups: normal, AGA, medication and EA group, 10 rats in each group. AGA model was established by induced monosodium urate (MSU) method, except the normal group. Tow days before AGA model was established, normal and AGA groups were lavaged with normal saline (20 ml/kg), medication group was lavaged with colchicine solution (20 ml/kg), EA(1.5-2 Hz, D.-D.wave, 9v; 1-3 rnA) was applied to "Sanyinjiao" (SP6), "jiexi" (ST41) and "Kunlun" (BL60) for 20 min, once daily;continuously for 9 days. Then observed the changes in dysfunction, and the content of TNF-α and IL-lβ detected by ELISA, the expression of TREM-l detected by immunohistochemistry and western blot.

RESULTSCompared to the normal group, the AGA group of the dysfunction index increased significantly (P<0.01), the content of TNF-α and IL-lβ increased significantly (P<0.05), the expression of TREM-l in synovial tissue increased significantly (P<0.05); the medication and EA groups compared to the AGA group, the dysfunction index decreased significantly (P<0.01), the content of TNF-α and IL-lβ decreased significantly (P<0.05), the expression of TREM-l in synovial tissue decreased significantly (P<0.05); there were not statistically significant between the medication and EA group (P>0.05).

CONCLUSIONEA treating AGA may be through down-regulating the expression of TREM -1 in synovial tissue.

Animals ; Ankle Joint ; metabolism ; pathology ; Arthritis, Gouty ; metabolism ; therapy ; Electroacupuncture ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Immunologic ; metabolism ; Synovial Membrane ; metabolism ; Triggering Receptor Expressed on Myeloid Cells-1 ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the effect of IPS-B2 on mouse peritoneal macrophages and the transcription of IL-1beta, IL-6, TNF-alpha and iNOS.

METHODELISA method and Griess method were used to detect the effect of mouse peritoneal macrophages produce cytokines IL-1beta, IL-6, TNF-alpha and cytotoxic effectors NO. The transcription of IL-1beta, IL-6, TNF-alpha and iNOS was detected by real-time RT-PCR method.

RESULTIPS-B2 could not promote mouse peritoneal macrophage production, but it could significantly improve the IL-1beta, IL-6, TNF-alpha content in mouse peritoneal macrophages culture supernatant, and increase the gene expression of IL-1beta, IL-6, TNF-alpha and iNOS.

CONCLUSIONIPS-B2 can enhance the ability of peritoneal macrophages to excrete bioactive substances and promote the transcription of bioactive substances to antitumor.

Agaricales ; chemistry ; Animals ; Gene Expression Regulation ; drug effects ; Interleukin-1beta ; biosynthesis ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; drug effects ; metabolism ; Mice ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Polymerase Chain Reaction ; Polysaccharides ; pharmacology ; RNA, Messenger ; biosynthesis ; Transcription, Genetic ; drug effects ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVEThis study aims to investigate the possible role and significance of soluble programmed death-1 (sPD-1)  /soluble programmed death ligand 1 (sPD-L1) in the immune pathogeneses of recurrent aphthous ulcer (RAU).

METHODSA total of 30 RAU cases (18 cases of minor RAU, 5 cases of major RAU, and 7 cases of herpetiform ulcers) were enrolled in this study. A total of 18 healthy people served as controls. Lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺, CD19⁺, and CD16⁺+56⁺) were investigated by flow cytometric analysis. Humoral immunity (IgG, IgA, IgM, C3, and C4) was explored by nephelometry immunoassay. The sPD-1 and sPD-L1 protein levels in the sera of RAU patients were investigated by enzyme-linked immunosorbent assay. The correlations of the sPD-1 and sPD-L1 protein levels with the immune status and clinical characteristics of the RAU patients were analyzed by SPSS 19.0.

RESULTSThe number of CD4+ T cells decreased and the levels of IgM antibodies increased in the RAU patients relative to those in the normal controls (P<0.05). The sPD-1 and sPD-L1 protein levels in the RAU patients were significantly higher than those in the control group (P<0.05). Meanwhile, the sPD-1 and sPD-L1 protein levels in the patients with minor and major RAU were significantly higher than those in the control group (P<0.05). By contrast, no significant difference was found in the patients with herpetiform RAU (P>0.05). Positive correlations were noted between the sPD-1 protein level and the CD19+ cell frequency or C4 level (r₁=0.389, P₁=0.034; r₂=0.382, P₂=0.037).

CONCLUSIONSCellular immune hypofunction and humoral immunity disorders were found in the RAU patients. The PD-1/PD-L1 signaling pathway, which might be influenced by the involvement of sPD-1 and sPD-L1 proteins to a certain extent, may play some roles in the immune pathogenesis of RAU.

B7-H1 Antigen ; Cell Count ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Programmed Cell Death 1 Receptor ; Signal Transduction ; Stomatitis, Aphthous