Objective To measure the expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis (EV), and to explore the relationship between cellular immune abnormality and EV pathogenesis. Methods Immunohistochemistry was used to measure the expressions of IL-10, IL-23 and CD86 in tissue samples from 10 patients with EV and 10 normal human controls. Results Three cytokines were observed in all the samples of EV, with the expression score ranging from 3 to 6 and expression intensity from moderate to high. However, of the control specimens, only 1 was positive for IL-10 with the expression score being 3, and expression intensity being moderate. Conclusion The pathogenesis of epidermodysplasia verruciformis may be correlated with the expression abnormality of some cytokines secreted by keratinocytes.

Objective To investigate the effects of a training program based on virtual reality on static and dynamic balance perfor-mance in patients with Parkinson's disease (PD). Methods From June, 2014 to June, 2016, 46 patients with PD were randomly divided into control group (n=23) and experimental group (n=23). The control group received routine balance training, while the experimental group re-ceived balance training of virtual reality, for six weeks. They were assessed with Unified Parkinson's Disease Rating Scale part 3 (UP-DRS3), Berg Balance Scale (BBS), TimedUp and GoTest (TUGT) and Hamilton Depression Scale (HAMD) before and after training. The envelope area, anteroposterior standard deviation (AP-SD), mediolateral standard deviation (ML-SD) of centre of pressure (COP) were also measured with posturography. Results The scores of BBS, TUGT and HAMD improved in both groups after training (t>2.657, P<0.05), and improved more in the experimental group than in the control group (t>2.426, P<0.05). The score of UPDRS3 and the parameters of pos-turography improved in the experimental group (t>2.626, P<0.05), and improved more than that in the control group (t>2.112, P<0.05). Con-clusion Virtual reality rehabilitation is more effective than routine balance training on the static and dynamic balance function in patients with PD, and may release their depression.

Objective Attenuated strains of Shigella are attractive live vaccine candidates for eliciting mucosal immune responses which is a suitable carrier for the prophylactic human papillomaviruses (HPV) vaccine development, To examine the potential of a live Shigella based prophylactic HPV vaccine, HPV16L1should be expressed in attenuated shigella strain. Methods A Shigella large invasive plasmid (icsA/virG) based prokaryotic expression plasmid pHS3199 was constructed. HPV16L1 gene was inserted into plasmid pHS3199 to form pHS3199-HPV16 L1 construct, and pHS3199-hpv16L1 was electroporated into a live attenuated shigella strain sh42. The expression of HPV16L1 protein was demonstrated by Western blotting with monoclonal antibody to HPV16L1, The genetic stability of recombinant strain sh42-HPV16 L1 was monitored by consecutive passage culture. Invasive ability of sh42-HPV16L1 was evaluated by Hela cell infection assay. Results HPV16 L1 protein can be expressed in recombinant strain sh42-HPV16 L1, and the protein stably expressed over 140 generations. The invasive ability of sh42-HPV16L1 was diminished dramatically compared to its parent strain, but not abolished completely. Conclusion HPV16L1 protein was constitutively expressed in the attenuated strain of shigella flexneri sh42, and maintained partial invasive ability. Our strategy may represent a promising vaccine candidate against genital HPV16 infection.

Objective:To investigate the effect of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in modulat-ing the effects of oral squamous cell carcinoma (OSCC) invasion. Methods:Real-time polymerase chain reaction was employed to de-tect the expression of MALAT1 in samples of OSCC post-radical resection, normal oral mucosa samples, and oral squamous cell lines. MALAT1-siRNA was transfected into TSCCa human tongue squamous cell carcinoma cell lines. Cell proliferation was determined by methyl-thiazolyl-tetrazolium reduction assay. Cell migration and invasive ability were evaluated by scratch test and transwell assay. The expression of proteins that regulated invasion and apoptosis were examined using Western blot assay. Immunofluorescence assay was used to detect changes in epithelial-mesenchymal transition (EMT)-associated proteins in the cells. Tumor-bearing nude mouse models were established by subcutaneous implantation of TSCCa cells. Immunohistochemistry was used to detect up-regulation of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2/9 (MMP-2/9). Results:MALAT1 expression was significantly higher in OSCC than in normal tissues (P<0.05). MALAT1 expression was inhibited by transfecting MALAT1-siRNA. After MALAT1 expres-sion was down-regulated in TSCCa cells, proliferation was inhibited and invasion was attenuated, showing significant differences com-pared with the cells transfected with scrambled siRNA and control cells (P<0.05). Expression of N-cadherin and MMP-2/9 were down-regulated in the cells after MALAT1 was knocked down. Tumor growth was significantly slower in the MALAT1-siRNA group than in the control groups. IHC indicated that PCNA and MMP-2/9 expression of tumor tissues were significantly inhibited in MALAT1-siR-NA group. Conclusion:MALAT1 is over-expressed in human OSCC. MALAT1 reduction can inhibit the proliferation and invasion of OSCC cells. Furthermore, MALAT1 may promote OSCC invasion and metastasis by modulating EMT.

Abstract: This study is to improve the affinity of scFv-AK404R against VEGFR2. The secondary mutational library was constructed by hydrophilic shuffling in CDR3 region of the heavy chain. VEGFR2-specific screening was performed by phage display technology and the protein of mutants was expressed in periplasm of E.coli HB2151 and purified by affinity chromatography. The affinity constant of scFvs was measured by competitive ELISA, and the structure of scFvs was analyzed by bioinformatics. The result showed that a library with 6.4x10(5) scFv members was established by electro-transformation. Two mutated clones with high absorbance value were isolated after screening. After purification by affinity chromatography, electrophoretically pure scFv proteins were obtained. The competitive ELISA showed that the affinities of WZ01 and WZ02 were three times higher than that of the parental AK404R, and bioinformatics analysis showed that the enlarged contact surface and fitted closely with KDR3 surface may be the reasons for improved affinity. These results suggest that introducing hydrophilic amino acids to the heavy chain CDR3 region is an effective approach to improve the affinity of scFv.
Amino Acid Sequence ; Antibody Affinity ; Chromatography, Affinity ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Hydrophobic and Hydrophilic Interactions ; Peptide Library ; Single-Chain Antibodies ; genetics ; immunology ; isolation & purification ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; metabolism

OBJECTIVETo study a new method for repair of facial depression and facial nerve defect after parotid carcinoma resection.

METHODS12 patients with parotid carcinoma and peripheral bone invasion were treated using facial nerve canal dissection and radical resection of the tumor, the parotid gland and the involved facial nerve and bone, including the mastoid, stylomastoid foramen, styloid process and the rear part of the mandible. A sternocleidomastoid muscle flap was elevated and transferred to repair the facial depression. The great annular nerve in the flap was anastomosed with the severed end of the facial nerve in the canal.

RESULTSThe depressed deformity of the parotid area was well corrected in 9 patients. The aesthetic results were compromised in 2 patients because of tumor recurrence and reoperation. The depressed deformity was not corrected in 1 patient because of infection. Postoperatively, the function of the facial nerve recovered to a normal level. The recovery time ranged from 12 to 20 weeks ,with an average of 16.3 weeks. The local control rate of tumor was improved.

CONCLUSIONSImmediate transplantation of the sternocleidomastoid muscle-great auricular nerve flap and facial nerve canal dissection in radical parotidectomy can repair the depressed deformity of the parotid area, restore facial nerve function,and decrease tumor recurrence. The method is an ideal operation with functional recovery.

Adult ; Cervical Plexus ; Facial Nerve ; transplantation ; Female ; Humans ; Male ; Middle Aged ; Neck Muscles ; transplantation ; Parotid Neoplasms ; surgery ; Surgical Flaps ; innervation ; Treatment Outcome

Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats. Methods The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy. Results After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P<0.05, P<0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P<0.01). The CaMKII immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMKII, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P<0.05, P<0.01). Conclusions The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMKII, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory.

The effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms were investigated. The rats were divided randomly into 5 groups (14 animals in each group): control, chronic hypoxia, chronic hypoxia with low (5 mg/kg body weight), middle (10 mg/kg body weight) and high (20 mg/kg body weight) concentrations of Rhizoma curcumae oil injection. The animals undergoing chronic hypoxia were exposed to hypoxia in a hypoxic chamber containing 10% O(2) and 5% CO(2) for 10 h/d, lasting 28 d. Morris water maze (MWM) test was used to obtain the scores of leaning and memory. The superoxide dismutase (SOD) activity and malonaldehyde (MDA) content were determined in the serum and hippocampus as well as [Ca(2+)](i) in the hippocampus. The expression of phosphorylated Ca(2+)/calmodulin-dependent protein kinase II (p-CaMKII) in the hippocampus was evaluated by using immunohistochemistry and Western blot. Compared with the control group, the chronic hypoxia group showed the following changes: (1) The escape latency to the hidden platform was remarkably prolonged (P<0.05); (2) The content of MDA and [Ca(2+)](i) were obviously higher, but the activity of SOD and the expression of p-CaMKII were significantly lower (P<0.05, P<0.01). Compared with the chronic hypoxia group, groups with Rhizoma curcumae oil injection had the following changes: (1) The escape latency to the hidden platform was remarkably shorter in 10, 20 mg/kg body weight groups (P<0.05); (2) The content of MDA and [Ca(2+)](i) were markedly decreased in 5, 10, 20 mg/kg body weight groups (P<0.05, P<0.01), but the activity of SOD in the serum and the expression of p-CaMKII were significantly higher in 10, 20 mg/kg body weight groups (P<0.05, P<0.01). The results showed that the capacity of learning and memory was degraded following chronic hypoxia. The decrease in MDA content and [Ca(2+)](i) and (or) the increase in SOD activity and p-CaMKII expression might participate in the enhancing effect on learning and memory induced by Rhizoma curcumae oil.
Animals ; Calcium ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Curcuma ; chemistry ; Hippocampus ; metabolism ; Hypoxia ; physiopathology ; Learning ; drug effects ; Malondialdehyde ; metabolism ; Memory ; drug effects ; Plant Oils ; pharmacology ; Rats ; Rhizome ; chemistry ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the expressions of CD1a and CD83 of Langerhans cells (LC) in the lesions of epidermodysplasia verruciformis (EV) patients.

METHODSWe used immunohistochemical method to detect the expressions of CD1a and CD83 in the lesions of 10 patients with EV lesions and in the skins of 10 normal subjects.

RESULTSNo CD83 + LCs was detected in all EV patients and normal controls, but CD1a + LC was found in all cases. The quantity of CD1a + LCs in the lesions of EV patients was significantly lower than that in the normal skin (P < 0.01); furthermore, the distribution of LCs in EV lesions was uneven.

CONCLUSIONThe functions of LCs may be inhibited in EV patients.

Antigens, CD ; biosynthesis ; genetics ; Antigens, CD1 ; biosynthesis ; genetics ; Epidermodysplasia Verruciformis ; immunology ; pathology ; Humans ; Langerhans Cells ; immunology ; Leukocyte Immunoglobulin-like Receptor B1 ; Receptors, Immunologic ; biosynthesis ; genetics ; Skin ; immunology ; pathology

OBJECTIVE To explore the neuro-protective effects of saffron (Crocus satius L.) on chronic focal cerebral ischemia in rats.METHODS SD rats were randomly divided into 6 groups:sham control group,MCAO group,edaravone group and saffron 30,100,300 mg·kg-1groups.Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).Saffron was administered orally by once daily from 2 h to 42 d after ischemia. At 42 d after cerebral ischemia, neurological deficit score, spontaneous activity test,elevated plus maze test,marble burying test and novel objective recognition test were used to evaluate the effects of saffron on the behevioural change. Infarct volume, survival neuron density, activated astrocyte number, and the thickness of glial scar were also detected. GFAP expression and inflammatory cytokine contents in ischemic peripheral region were detected by Western blot and ELISA,separately.RESULTS Saffron(100,300 mg·kg-1)improved the body weight decrease, neurological deficit and spontaneous activity. Saffron (30-300 mg·kg- 1) increased the traveled distance ratio and total time in open arm, decreased the buried marble number, which indicated that saffron could ameliorate anxiety- and depression-like behaviors. Saffron (100, 300 mg·kg-1)improved the learning and memory function,which manifested by increased discrimination ratio(DR)and discrim-ination index (DI) in T2test. The results of toluidine blue found saffron treatment (100, 300 mg·kg-1) decreased the infarct volume and increased the neuron density in cortex and hippocampal.The activated astrocyte number,the thickness of glial scar and GFAP expression in ischemic peripheral region decreased after saffron. Saffron (100, 300 mg·kg-1) decreased the contents of IL-6 and IL-1β, increased the content of IL-10 in ischemic peripheral region.CONCLUSION Saffron exerted neuro-protective effects on chronic focal cerebral ischemia,which could be related with inhibiting the activation of astrocyte and glial scar,following with the decrease of inflammatory reaction.