With homology cloning approaches coupling with RACE (rapid-amplification of cDNA ends) techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF (named as PnPR10-1 with the Accession No. KJ741402 in GenBank) was detected from the cloned sequence with full-length of cDNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of PnPR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Genes, Plant ; Glycoside Hydrolases ; genetics ; Molecular Sequence Data ; Open Reading Frames ; Panax notoginseng ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plant Roots

ObjectiveTo discuss the relationships among spastic paralysis and motor function, balance function and other clinical variables after stroke.MethodsAssessed the clinical spastic index of hemiplegic lower limbs with CSI scale, motor function with Fugl-Meyer Assessment, and balance function with Berg Equilibrium Scale, respectively. And then analyzed the relationship between clinical spasm index with motor function, balance function of the hemiplegic lower limbs with Pearson correlation analysis. Finally, ascertained which was the most important factor affecting the clinical spasm index of spastic lower limbs with stepwise regression analysis.ResultsThere is negative correlation between clinical spasm index with motor function and balance function, respectively. Tendon reflex, muscular tension and clonus are the deciding factors to motor function(P<0.05) and balance function(P<0.05) of the hemiplegic lower limbs.ConclusionThe clinical spasm index of hemiplegic limbs can affect the recovery of motor function and balance function significantly. So, assessing and improving the clinical spasm index of paralytic lower limbs can optimize the rehabilitation program to stroke patients.

OBJECTIVETo investigate the protective effect of Danxuetong injection (DXT, a combination of Danshen and Xueshuantong injections) against testicular ischemia-reperfusion injury following testis torsion/detorsion in rats.

METHODSThirty-two 4-week-old healthy male SD rats were randomly divided into four groups of equal number: sham operation, normal saline, single DXT injection, and successive DXT injection. The rat models of testicular ischemia-reperfusion injury were established by 2-hour 720-degree torsion/detorsion of the unilateral testis. At 6 weeks after modeling, the rats were killed and their testes were harvested for measure- ment of testicular coefficients, sperm counts, sperm motility, and the levels of total anti-oxidative capacity (T-AOC) , superoxide dismutase (SOD) , nitric oxide synthase (NOS) , and malondialdehyde ( MDA) in the testis tissue.

RESULTSCompared with the rats of the normal saline group, those of the single DXT injection and successive DXT injection groups showed significant increases in the testicular coefficient (0.11 ± 0.03 vs 0.35 ± 0.04 and 0.40 ± 0.06, P < 0.05), sperm count ([0.46 ± 0.10] vs [1.44 ± 0.50] and [3.00 ± 1.28] x10(9)/ml, P < 0.05), sperm motility ([13.63 ± 14.04] vs [39.63 ± 5.04] and [76.31 ± 3.67]%, P < 0.05), the activity of SOD (72.76 ± 5.58 vs 116.25 ± 8.83 and 133.20 ± 13.84, P < 0.05), and the level of T-AOC (5.58 ± 1.07 vs 13.34 ± 5.81 and 19.21 ± 5.69, P < 0.05), but a remarkable decrease in the content of MDA (42.38 ± 8.94 vs 20.94 ± 5.65 and 15.02 ± 1.03, P < 0. 05) in the injured testes.

CONCLUSIONDXT can effectively rid the testis tissue of oxygen free radicals, improve sperm count and motility by antioxidation, and protect the testis tissue of prepubertal rats against testicular ischemia-reperfusion injury after testis torsion/detorsion. It also has a protective effect on the contralateral testis, and successive injection has a better effect than single injection of DXT.

Animals ; Antioxidants ; therapeutic use ; Drug Therapy, Combination ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Spermatic Cord Torsion ; complications ; therapy ; Superoxide Dismutase ; metabolism ; Testis ; blood supply ; metabolism

The mediastinal lymph node tuberculous abscesses (MLNTAs) are secondary to mediastinal tuberculous lymphadenitis.Surgical excision is often required when cold abscesses form.This study was aimed to examine video-assisted thoracoscopic surgery (VATS) for the treatment of MLNTA.Clinical data of 16 MLNTA patients who were treated in our hospital between December 1,2013 and December 1,2015 were retrospectively analyzed.All of the patients underwent the radical debridement and drainage of abscesses,and intrathoracic lesions were removed by VATS.They were also administered the intensified anti-tuberculosis treatment (ATT),and engaged in normal physical activity and follow-up for 3 to 6 months.The results showed that VATS was successfully attempted in all of the 16 MLNTA patients and they all had good recovery.Two patients developed complications after surgery,with one patient developing recurrent laryngeal nerve injury,and the other reporting poor wound healing.It was concluded that VATS is easy to perform,and safe,and has high rates of success and relatively few side-effects when used to treat MLNTA.

Objective To observe the effects of repeated Botulinum toxin type A (BTX-A) injection on lower limb spasticity after stroke.Methods 180 cases with lower limb spasticity after stroke were divided into the treatment group (n=90) and the control group (n=90). The treatment group was treated with BTX-A injection twice in the spastic muscles at interval of 3~6 months, while both the treatment group and the control group accepted the rehabilitation based on the neurodevelopmental therapy. They were assessed with modified Ashworth Scale (MAS), Fugl-Meyer Lower Limb Assessment (FMAL), Berg Balance Scale (BBS), modified Barthel Index (MBI) before each injection, and 3 d, 7 d, 1 month, 3 months after each injection or the same time for the controls. Results There was significant difference in scores of MAS, FMAL, BBS, MBI for the treatment group among before and 3 d, 7 d, 1 month after each injection (P<0.05), but not significant between 2 injections (P<0.05). There was significant difference in scores of all the assessment between the treatment and control group at the same time (P<0.01). Conclusion Repeated intramuscular injection of BTX-A can reduce the spasticity of lower limb after stroke.

OBJECTIVE: To detect the serum level of a novel autoantibody, anti-tubulin-α-1C, in patients with systemic sclerosis (SSc) and to investigate its clinical significance. METHODS: Anti-tubulin-α-1C antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) in 62 patients with SSc, 38 systemic lupus erythematosus (SLE), 24 primary Sjögren's syndrome (pSS) patients, and 30 healthy controls (HCs). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin A(IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), C3, C4, rheumatoid factor (RF), antinuclear antibody(ANA), anti-centromere antibodies(ACA), anticardiolipin (aCL), anti-dsDNA antibody, anti-Sm antibody, anti-RNP antibody, anti-Scl-70 antibody, anti-Ro52 antibody, anti-SSA antibody, anti-SSB antibody, centromere protein A(CENP-A), centromere protein B (CENP-B) were measured by standard laboratory techniques. Raynaud's phenomenon and modified Rodnan skin score(MRSS) were recorded to evaluate the disease status of SSc. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman rank correlation were used for statistical analyses. RESULTS: The serum anti-tubulin-α-1C antibody concentration in SSc group was 81.24±34.38, the serum anti-tubulin-α-1C antibody concentration in SLE group was 87.84±38.52, the serum anti-tubulin-α-1C antibody concentration in pSS group was 59.79±25.24, and the serum anti-tubulin-α-1C antibody concentration in healthy group was 39.37±18.7. Multivariate analysis revealed that anti-tubulin-α-1C antibody levels were significantly increased in the SSc and SLE patients. The expression level of anti-tubulin-α-1C antibody in SSc was higher compared with the pSS group and the health control group (P < 0.01). Further analysis demonstrated that the elevated anti-tubulin-α-1C antibody were correlated with the SSc inflammation and disease activity markers ESR(r=0.313, P=0.019), The levels of anti-tubulin-α-1C antibody were also significantly correlated with MRSS(r=0.636, P < 0.01). The best cut-off value for the diagnose of SSc was 76.77 as mean+2SD value. The proportion of Raynaud's phenomenon was higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than that in anti-tubulin-α-1C autoantibody negative group(71.4% vs. 37.5%, P=0.039). The proportions of anti-Scl-70 antibody, anti-CENP antibody and anti-cardiolipin antibody were higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than in the anti-tubulin-α-1C autoantibody negative group (37.9% vs. 15.2%, 34.5% vs. 12.1%, 13.8 vs. 0, respectively, all P < 0.05). CONCLUSION Based on this explorative stu-dy, the level of anti-tubulin-α-1C antibody increased in the serum of the patients with SSc. There were correlations between anti-tubulin-α-1C autoantibody and clinical and laboratory indicators of the SSc patients. It may become a novel biomarker indicative of active SSc and could be applied in future clinical practice.
Antibodies, Antinuclear ; Autoantibodies ; Humans ; Lupus Erythematosus, Systemic ; Scleroderma, Systemic ; Sjogren's Syndrome

BACKGROUNDThe mechanisms underlying postoperative pain remain unclear. Neurotransmitters of excitatory and inhibitory amino acids play an important role in the transmission and modulation of pain in the spinal dorsal horn. This study aimed to investigate the changes of release of excitatory and inhibitory amino acids in the spinal cord during postoperative pain and to provide a novel theoretical basis for postoperative pain management.

METHODSLoop microdialysis catheters were implanted subarachnoidally via the atlanto-occipital membrane in 16 healthy Sprague-Dawley rats. All rats without neural deficits were divided into two groups, Group A and Group B, following 5 days of recovery. The tubes for microdialysis were connected and 25 microl microdialysate sample for baseline value was collected after one-hour washout in each rat. A plantar incision in the right hind paws of rats in Group A were performed under 1.2% isoflurane. All rats in Group B were only anesthetized by 1.2% isoflurane for the same duration. The microdialysate samples were collected at 3 hours, 1 day, 2 days and 3 days after the incision (or isoflurane anesthesia in Group B) in both groups. The cumulative pain scores were also assessed at the above time-points. The amino acids in the microdialysate samples were tested using high performance liquid chromatography.

RESULTSWithin Group A, the release of aspartate and glutamate at 3 hours after the incision was significantly higher than the baseline values and the release of glycine at 1 day after the incision significantly increased compared with the baseline values (P < 0.01). Within Group B, the release of neurotransmitters at each time point had no significant difference compared with the baseline values (P > 0.05). The release of aspartate and glutamate at 3 hours after the incision in Group A was significantly higher than that in Group B (P < 0.01). The release of glycine at 1 day after the incision in Group A significantly increased compared with Group B (P < 0.01). The cumulative pain scores at 3 hours, 1 day and 2 days after the incision in Group A were significantly higher than those in Group B (P < 0.01).

CONCLUSIONSThe release of the excitatory amino acids occurs in the early phase of postoperative pain and might not be involved in the maintenance of pain in a rat model of incision pain. The release of inhibitory glycine lagged behind the excitatory amino acids. The implication of inhibitory glycine release remained to be established further.

Animals ; Aspartic Acid ; secretion ; Excitatory Amino Acids ; cerebrospinal fluid ; secretion ; Glutamic Acid ; secretion ; Glycine ; secretion ; Male ; Microdialysis ; Neurotransmitter Agents ; secretion ; Pain, Postoperative ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; secretion

OBJECTIVETo study the role of herpes simplex virus type 1 ( HSV-1 ) in facial paralysis by developing an experimental animal model of viral facial paralysis.

METHODSBoth sides of posterior auricular branch of facial nerve were anatomies and incised in 66 mice. The HSV-1 was inoculated into right ear branch and fetal bovine serum was inoculated into left ear branch as control. The symmetry of mouse face was observed and scored. The temporal bones were serially sectioned and stained with hematoxylin and eosin. The extratemporal facial nerves were stained with osmium tetroxide. HSV-1 DNA in bilateral facial nerve, brain stem, trigeminal ganglion and spinal cord was detected by the polymerase chain reaction.

RESULTSTwenty-eight (42. 42%) mice developed right facial paralysis between 2 and 5 days after inoculation. Continuing 3-6 days, the facial paralysis recovered spontaneously. Thirty-eight mice had no signs of facial paralysis. Compared with the left, nerve swelling, inflammatory cell infiltration were manifested in right temporal facial nerve of paralyzed mice. The ratio of the cross-sectional area of the facial nerve to the facial canal ( FN/FC ) was significantly higher than that on the control side (P < 0.01). Demyelinated nerve fibers were seen in the right extratemporal facial nerve. Not only in paralyzed mice, but also in non-paralyzed mice, HSV DNA was detected in some nerve tissues.

CONCLUSIONSInoculating HSV-1 into posterior auricular branch of facial nerve can produce an acute and transient facial paralysis in mice. The possible pathophysiologic mechanism of the facial paralysis is viral invasion and transportation from distal branch to main trunk. Then the viral facial neuritis causes facial paralysis.

Animals ; Disease Models, Animal ; Facial Nerve ; virology ; Facial Nerve Diseases ; virology ; Female ; Herpes Simplex ; physiopathology ; Herpesvirus 1, Human ; Mice ; Mice, Inbred BALB C

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Glycyrrhizic Acid ; pharmacology ; Humans ; Lipopolysaccharides ; toxicity ; Liver ; drug effects ; pathology

OBJECTIVETo investigate the expressions of myogenic markers MyoD, myogenin,and desmin in skeletal muscle differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs).

METHODSMyogenic markers MyoD, myogenin,and desmin of hBM-MSCs cultured in vitro were detected by immunofluorescence and RT-PCR. A total of 21 8-to-10 week-old immunosuppressed mdx mice were transplanted with 1x107 passage 5 of hBM-MSCs. The mice were euthanized 2-24 weeks after transplantation,and gastrocnemius muscle were analyzed for human MyoD, myogenin,desmin,and dystrophin (Dys) expressions by immunohistochemistry and RT-PCR.

RESULTSThe numbers of MyoD-,myogenin-,and desmin-positive cells per 100 hBM-MSCs were 23.5∓5.3, 30.7∓6.2, and 28.4∓5.7, respectively. MyoD, myogenin, and desmin mRNA was observed in passage 5 of hBM-MSCs. After two weeks of hBM-MSCs transplantation,a small number of MyoD-and myogenin-positive cells were observed in skeletal muscle of mdx mice,and desmin-positive cells were observed 4 weeks after transplantation. Expressions of MyoD and myogenin were detected in the muscle of mdx mice 2-4 weeks after hBM-MSCs transplantation, which reached a peak 12-16 weeks later. Desmin was expressed in the muscle of mdx mice 4-8 weeks after transplantation,with much more expression after 16 weeks of transplantation. A small number of Dys-positive cell and Dys mRNA expression were presented in the muscle of mdx mice 4 and 8 weeks after hBM-MSCs transplantation,respectively. The expression of Dys in the muscle of mdx mice increased gradually after transplantation.

CONCLUSIONhBM-MSCs have the potential of myogenic differentiation in vitro and contribute to myogenic conversion in xenogeneic animal,during which the up-regulation of MyoD and myogenin expressions may play an important role.

Animals ; Biomarkers ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Desmin ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred mdx ; Muscle, Skeletal ; cytology ; metabolism ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Up-Regulation