BACKGROUNDCD44v6 plays an important role in invasion and metastasis of tumor, Livin has anti-apoptotic effects. The present study aimed to explore the expression and clinical significance of CD44v6 and Livin in gastric cancer tissue.

METHODSStreptavidin-peroxidase linked immunohistochemical method was used to determine the expression of CD44v6 and Livin in gastric cancer tissue and adjacent normal gastric tissues from 59 patients with histopathologically confirmed gastric cancer, and in gastric tissue specimens of 15 patients with gastric polyps, and 15 patients with chronic non-atrophic gastritis. The chi-square test was used for comparison of the relevant factors, Spearman's rank correlation test was applied for relationship among positive expression of the proteins.

RESULTSThe expresion of CD44v6 was positive in 64.4% of the gastric cancer patients; 5.1%, 0 and 13.3% in specimens of normal tissues adjacent to the cancer tissues, in gastric tissue specimens of patients with gastric polyps, and patients with chronic non-atrophic gastritis, respectively. The expression of Livin was positive in 52.5% of the gastric cancer tissues, 6.8%, 0 and 6.7% in the adjacent normal gastric tissue, specimens of patients with gastric polyps and chronic non-atrophic gastritis, respectively. The expression of CD44v6 was significantly correlated with the depth of invasion, the degree of differentiation, and lymphnode metastasis of gastric cancer (P < 0.05). The positive expression rate of Livin protein was also significantly correlated with degree of differentiation of gastric cancer cells and metastasis to lymphnodes (P < 0.05), but not correlated with the depth of invasion and pathological types (P > 0.05). The expression of CD44v6 and Livin in the gastric cancer tissue was positively correlated (r(s) = 0.286, P = 0.028).

CONCLUSIONSThe increased expression of CD44v6 and Livin in gastric cancer tissue may be closely related with development and progression of gastric cancer. CD44v6 and Livin may be new biological markers of gastric cancer.

Adaptor Proteins, Signal Transducing ; analysis ; physiology ; Aged ; Female ; Humans ; Hyaluronan Receptors ; analysis ; physiology ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; analysis ; physiology ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; physiology ; Stomach Neoplasms ; chemistry ; metabolism ; pathology

Objective:To investigate the analgesic effect of neurotropin (NTP) on bone cancer pain (BCP) and its preliminary mechanisms in rats.Methods:(1) According to the random number table method, 72 Sprague-Dawley (SD) female rats were randomly and equally divided into 6 groups: normal control group, sham-operated group, BCP model group, BCP+low-dose NTP group, BCP+medium-dose NTP group, and BCP+high-dose NTP group ( n=12). The SHZ-88 breast cancer cells were inoculated into the tibias of rats in the latter 4 groups to establish BCP models. After 15-21 d of modeling, the rats of the latter 3 groups were intraperitoneally administered with 1.2, 2.4 and 3.6 unit NTP, respectively, once per d for 7 consecutive d. The mechanical withdrawal threshold (MWT) changes were measured in each group before BCP and 5, 7, 10, 14, 17, and 21 d after BCP. The number of 5-hydroxytryptamine 7 (5-HT7) positive cells and 5-HT7 protein expression in the ventrolateral periaqueductal gray (vlPAG) were detected by immunohistochemistry and Western blotting, respectively. (2) Twenty-four rats 21 d after BCP modeling were randomly divided into BCP group, BCP+high-dose NTP group (BCP rats with intraperitoneal injection of 3.6 unit NTP), BCP+NTP+SB-269970 group (BCP rats with pretreatment of specific 5-HT7 receptor antagonist SB-269970 in the vlPAG for 30 min, and then, with intraperitoneal injection of 3.6 unit NTP, n=8). The MWT changes were measured in each group before NTP and 15, 30, 45, 60 min after NTP. Results:(1) Seventeen and 21 d after modeling, the MWT values of the modeled hind limb of rats in BCP+low-dose NTP group, BCP+medium-dose NTP group, and BCP+high-dose NTP group were significantly higher than those in BCP group; those in the BCP+high-dose NTP group were significantly higher than those in BCP+low-dose NTP group and BCP+medium-dose NTP group; those in BCP+medium-dose NTP group were statistically higher than those in the BCP+low-dose NTP group ( P<0.05). Twenty-one d after modeling, the number of 5-HT7 receptor positive cells and protein expression in the vlPAG of rats in the BCP+low-dose NTP group, BCP+medium-dose NTP group, and BCP+high-dose NTP group were significantly larger/higher than those in BCP group; those in the BCP+high-dose NTP group were significantly larger/higher than those in BCP+low-dose NTP group and BCP+medium-dose NTP group; those in BCP+medium-dose NTP group were statistically larger/higher than those in the BCP+low-dose NTP group ( P<0.05). (2) At 15, 30, 45 and 60 min after injection of NTP, the MWT values of the modeled hind limb of rats in the BCP+high-dose NTP group and BCP+NTP+SB-269970 group increased gradually, enjoying statistical significance as compared with those in the BCP group at the same time point ( P<0.05); however, the MWT values of the BCP+NTP+SB-269970 group were significantly lower as compared with those in the BCP+high-dose NTP group at the same time point ( P<0.05). Conclusion:The activation of 5-HT7 receptor in the vlPAG is involved in the analgesic effect of NTP on BCP in rats.

BACKGROUND: Chinese medicine is effective for preventing and treating glucocorticoid-induced osteoporosis, however, the underlying mechanism remains unclear. DKK1, an inhibitor of Wnt/β-catenin signaling pathway, can be up-regulated by glucocorticoid. Thereafter, DKK1 is an important target in the prevention and treatment of osteoporosis. OBJECTIVE: To explore the regulatory effect of Zuogui pill on DKK1 in the prevention and treatment of glucocorticoid-induced osteoporosis. METHODS: Eighteen three-month-old female Sprague-Dawley rats were randomly divided into three groups: control group, model group and Zuogui pill group. Rats in the model and Zuogui pill groups received the subcutaneous injection of dexamethasone to establish the model of glucocorticoid-induced osteoporosis. The Zuogui pill group rats were administrated Zuogui pill extracts, and the control rats were given the same volume of normal saline. At 1 month after modeling, the lumbar vertebrae were removed to test the bone mass and microstructures by micro-CT scanning. The biomechanical properties were detected by compression test. The mRNA expression levels of DKK1, Runx2 and CTSK were determined by Qpcr. The serum alkaline phosphatase activity was tested. RESULTS AND CONCLUSION: Compared with the control group, in the model group, the volumetric bone mineral density, trabecular bone volume fraction, trabecular number, and trabecular thickness were significantly decreased (P ＜ 0.05), the trabecular separation and structure model index were significantly increased (P ＜ 0.05). The serum alkaline phosphatase activity was on a decline. The mRNA expression level of DKK1 showed a significant up-regulation (P ＜ 0.05). The mRNA expression level of Runx2 showed a down-regulated trend while mRNA expression level of CTSK showed an up-regulated trend. Compared with the model group, the Zuogui pill group showed significantly enhanced volumetric bone mineral density, trabecular bone volume fraction, and trabecular number (P ＜ 0.05); the structure model index was significantly decreased (P ＜ 0.05); the trabecular separation was reduced; the serum alkaline phosphatase activity was enhanced; the mRNA expression level of DKK1 showed a significant down-regulation (P ＜ 0.05); the mRNA expression level of Runx2 showed an up-regulated trend while mRNA expression level of CTSK showed a down-regulated trend. The vertebral compressive strength in the Zuogui pill group was significantly higher than that in the model group (P＜0.05). In summary, Zuogui pill prevents and treats glucocorticoid-induced osteoporosis possibly through the down-regulation of mRNA expression of DKK1.

BACKGROUNDAttention-deficit hyperactivity disorder (ADHD) is the most common mental and behavioral disorder in school-aged children. This study evaluated the effect of osmotic-release oral system (OROS) methylphenidate (MPH) on cognitive function and academic performance of Chinese school-aged children with ADHD.

METHODSThis 12-week, prospective, multicenter, open-label, self-controlled study enrolled 153 Chinese school-aged children with ADHD and 41 non-ADHD children. Children with ADHD were treated with once-daily OROS-MPH (18 mg, 36 mg, or 54 mg). The primary endpoints were Inattention/Overactivity (I/O) with Aggression Conners Behavior Rating Scale (IOWA) and Digit Span Test at week 12 compared with baseline. Secondary endpoints included opposition/defiant (O/D) subscale of IOWA, Clinical Global Impression (CGI), Coding Test, Stroop Color-word Test, Wisconsin Card Sorting Test (WCST), academic performance on teacher-rated school examinations, and safety at week 12 compared with baseline. Both non-ADHD and ADHD children received the same frequency of cognitive operational test to avoid the possible bias caused by training.

RESULTSA total of 128 patients were evaluated with cognitive assessments. The OROS-MPH treatment significantly improved IOWA Conners I/O subscale scores at week 12 (3.8 ± 2.3) versus baseline (10.0 ± 2.4; P < 0.0001). Digit Span Test scores improved significantly (P < 0.0001) with a high remission rate (81.1%) at week 12 versus baseline. A significant (P < 0.0001) improvement was observed in O/D subscale of IOWA, CGI, Coding Test, Stroop Color-word Test, WCST, and academic performance at week 12 versus baseline. Very few practice-related improvements were noticed in the non-ADHD group at week 12 compared with baseline. No serious adverse events and deaths were reported during the study.

CONCLUSIONSThe OROS-MPH treatment effectively controlled symptoms of ADHD and significantly improved academic performance and cognitive function of Chinese school-aged children with ADHD. The treatment was found to be safe and generally well-tolerated over 12 weeks.

TRIAL REGISTRATIONClinicalTrials.gov, NCT01933880; http://clinicaltrials.gov/ct2/show/NCT01933880?term=CONCERTAATT4099&rank=1.

Administration, Oral ; Attention Deficit Disorder with Hyperactivity ; drug therapy ; physiopathology ; Child ; Cognition ; drug effects ; Female ; Humans ; Male ; Methylphenidate ; administration & dosage ; adverse effects ; therapeutic use ; Neuropsychological Tests ; Prospective Studies ; Treatment Outcome

Adolescent ; Child ; Child, Preschool ; Female ; Hematopoiesis ; Humans ; Infant ; Leukocyte Count ; Male ; Neoplasms ; blood ; mortality ; surgery ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous

OBJECTIVETo observe the effect of recombinant human erythropoietin (rhuEPO) on p-Akt and caspase-9 expressions in the hippocampus of rats with status epilepticus (SE) and explore the neuroprotective mechanism of rhuEPO.

METHODSAdult male SD rats were randomized into control, PTZ, rHuEPO, LY294002 group, and DMSO groups and treated with normal saline (NS), PTZ, PTZ+rHuEPO, PTZ+LY294002+rHuEPO, and PTZ+DMSO+rHuEPO, respectively. The behavioral and electroencephalogram (EEG) changes of the rats were recorded, and the expressions of p-Akt and caspase-9 were detected using immunohistochemistry. The hippocampal expression of caspase-9 mRNA was detected using RT-PCR, and the expressions of Akt and p-Akt proteins were determined with Western blotting.

RESULTSThe p-Akt-positive cell and p-Akt protein expression increased significantly while the caspase-9-positive cell and caspase-9 mRNA expression decreased in rHuEPO group as compared with those in PTZ group (P<0.05). LY294002 treatment prior to rHuEPO injection significantly abolished the effects of rHuEPO on caspase-9 and p-Akt immunohistochemical positivity and caspase-9 mRNA and p-Akt protein expressions (P<0.05).

CONCLUSIONAdministration of rHuEPO activates the PI3K/Akt signaling pathway in SE rats and increases the expression of p-Akt protein to regulate the expression of caspase-9, a regulatory factor of the mitochondrial-dependent apoptotic pathway, and therefore provides anti-apoptotic and neuroprotective effects.

Animals ; Caspase 9 ; genetics ; metabolism ; Erythropoietin ; therapeutic use ; Hippocampus ; metabolism ; Male ; Neuroprotective Agents ; therapeutic use ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; Status Epilepticus ; drug therapy ; metabolism

BACKGROUNDA monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).

METHODSA phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.

RESULTSSix positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07 x 10(6) individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.

CONCLUSIONSThe results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

Animals ; Antibodies, Helminth ; immunology ; Antibodies, Monoclonal ; immunology ; Antigens, Helminth ; blood ; Base Sequence ; Immunoglobulin Fragments ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Peptide Library ; Rabbits ; Schistosomiasis japonica ; diagnosis ; Sensitivity and Specificity ; Serologic Tests

OBJECTIVETo observe the treating effect of collage-heparin sulfate after the 10 mm rat sciatic nerve defect was bridged by it.

METHODSA new kind of nervous tissue engineering scaffold was produced by freeze-drying technique from collagen-heparin sulfate. Thirty-two SD rats were randomly divided into A, B, C and D groups. Sciatic nerve defect in group A was bridged by collagen-heparin sulfate. In group B, sciatic nerve was bridged by auto-nerve transplantation. Group C was the blank control group. Animals in group D were normal. And 10 mm sciatic nerve defect was bridged in the experiment. Thirty-six weeks after the operation, the experimental animals were detected by HRP labeled retrograde trace, HE staining, toluidine staining, silvering staining, S100, GAP-43 and NF immunohistological staining, MBP immunofluorescence staining and transmission electron microscope to observe the nerve regeneration inducing effect of this new scaffold.

RESULTSNine months after operation, the collage-heparin sulfate scaffold was replaced by newly regenerated nerve. The number of HRP labeled spinal cord anterior horn cells and the area of sensation nerve fiber at the posterior horn were similar with that was repaired by auto-nerve. GAP-43, NF and S100 labeled regenerated nerve fiber had passed the total scaffold and entered the distal terminal. The regenerated nerve fibers were paralleled, lineage arranged, coincide with the prearranged regenerating "channel" in the collagen-heparin sulfate scaffold. MBP immunofluorescence staining also proved that the newly regenerated nerve fiber could be ensheathed. In the experimental group, the area of myelinated nerve fiber and the thickness of the myelin sheath had no obvious difference with that of the group repaired by auto-nerve, except that the density of the regenerated myelinated sheath fiber was lower than that of the control group.

CONCLUSIONNervous tissue engineering scaffold produced by collagen-heparin sulfate can guide the regeneration of nerve fibers. The nerve function recovers fine. This kind of material has great application potential.

Animals ; Biocompatible Materials ; Heparitin Sulfate ; Male ; Prosthesis Implantation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; pathology ; surgery ; Sulfuric Acid Esters ; Tissue Engineering ; methods

OBJECTIVETo evaluate the gene-chip detecting rifaman-resistance Mycobacterium tuberculosis applied in TB diagnosis and drug-resistant detection.

METHODSMycobacterium tuberculosis and rifaman-resistant strains among 35 rifaman-resistance isolated strains and 102 sputa specimens from TB patients, 27 sputa specimens from other patients were examined the gene-chips. Results obtained were compared with sputum examination, bacteriological culture and standard drug susceptibility test of Mycobacterium tuberculosis.

RESULTSThirty-five rifaman-resistance strains were detected by gene-chips and 33 were identified as rifaman-resistance strains and the concordance with the traditional drug susceptibility test of Mycobacterium tuberculosis was 94.29%. Twenty-seven sputa specimens from other patients were examined Mycobacterium tuberculosis by the gene-chips, 2 were positive, the detection specialty was 92.59%. Using three methods detecting Mycobacterium tuberculosis among 102 sputa specimens the positive rate respectively was, sputum examination 35.29% (36/102), bacteriological culture 28.43% (29/102), gene-chip 77.45% (79/102). Among 102 sputa specimens only 29 examined Mycobacterium tuberculosis by the traditional drug susceptibility test and 8 were rifaman-resistant strains. While using gene-chip, there were 20 among 102 sputa specimens identified as rifaman-resistance strains. Among total 55 rifaman-resistance strains detected by the gene-chips, the most frequent mutations were those associated with codon 531 (23 of 55; 41.8%), 526 (15 of 55; 27.27%) and 516 (9 of 55; 16.36%).

CONCLUSIONResults showed that this was a rapid, simple and highly specific method when using gene-chip to detect Mycobacterium tuberculosis and rifaman-resistant strains.

China ; epidemiology ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; genetics ; Female ; Humans ; Male ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; Point Mutation ; Rifampin ; pharmacology ; Sensitivity and Specificity ; Sputum ; microbiology ; Tuberculosis, Multidrug-Resistant ; epidemiology ; microbiology ; Tuberculosis, Pulmonary ; epidemiology ; microbiology

OBJECTIVETo prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovirus 71.

METHODSTwo peptides, SP55 and SP70, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to establish the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test.

RESULTSTwo high titered anti-VP1 antibodies secreted by the hybridoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1:16 respectively.

CONCLUSIONTwo high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; Antibodies, Viral ; analysis ; immunology ; Capsid Proteins ; analysis ; immunology ; Enterovirus ; chemistry ; immunology ; Enterovirus Infections ; diagnosis ; immunology ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neutralization Tests