Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P

OBJECTIVETo observe the effect of Xinfeng Capsule (XFC) on ankylosing spondylitis (AS) patients' symptoms and signs, serum immunoglobulin levels, peripheral blood lymphocyte autophagy protein, autophagy gene, and to explore its mechanism.

METHODSTotally 59 AS patients were assigned to the treatment group (39 cases) and the control group (20 cases) according to random digit table. Patients in the treatment group received XFC, 0.5 g each pill, three pills each time, 3 times per day, while those in the control group received sulfasalazine (SASP), 0.25 g per tablet, 4 tablets each time, twice per day. Three months consisted of one therapeutic course. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) were statistically calculated. Serum immunoglobulins (IgG1, IgG2, IgG3, IgG4, IgA , SIgA, and IgM) were detected using ELISA. Changes of Beclin1, LC3-II, phosphatidylinositol 3-kinase (PI3K), Akt, the mammalian target of rapamycin (mTOR) were detected using Western blot. Serum autophagy related genes such as Atg1, Atg5, Atg12, Atg13, and Atg17 were detected using the polymerase chain reaction (PCR). The correlation between immunoglobulin subtypes and autophagy gene in AS patients using Spearman correlation.

RESULTSCompared with before treatment, BASDAI, IgG1, lgG3, and IgA decreased (P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.01); ATG1, ATG12, ATG13, and ATG17 mRNA expressions decreased, ATG5 mRNA expression increased (P < 0.01) in the treatment group. But BASDAI, IgG1, and IgA levels decreased (P < 0.05, P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.05); ATG1 and ATG13 mRNA expressions decreased (P < 0.05, P < 0.01) in the control group. Compared with the control group, BASDAI, IgG1, and IgA levels decreased (P < 0.05); PI3K, Akt, mTOR protein expressions decreased (P < 0.01); ATG12 and ATG17 mRNA expression decreased, ATG5 mRNA expression increased (P < 0.01) in the XFC group. Correlation analysis showed AS patients' IgG1, IgG2, IgG3, IgA, SIgA, IgM had negative correlation with ATG17; IgG4 and ATG17 were positively correlated (P < 0.05, P < 0.01).

CONCLUSIONXFC could elevate clinical efficacy of AS patients and enhance their autophagy, which might be achieved by acting on PI3K/Akt/mTOR signal, affecting autophagy gene and autophagy protein expression, taking part in the regulation of proliferation and differentiation of lymphocyte B, and strengthen humoral immunity.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; Beclin-1 ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Lymphocytes ; drug effects ; Membrane Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Spondylitis, Ankylosing ; drug therapy ; Sulfasalazine ; therapeutic use ; TOR Serine-Threonine Kinases ; metabolism

OBJECTIVETo study the expression of vascular endothelial growth factor-C (VEGF-C) in oral squamous cell carcinoma (OSCC) and its relation to angiogenesis, lymphangiogenesis, as well as lymph node metastasis.

METHODSSixty-seven archival specimens from patients with oral squamous cell carcinoma were investigated, whose clinicopathologic data were completely conserved. Immunohistochemical staining was performed to detect the expression of VEGF-C, microvessel density (MVD), lymphatic vessel density (LVD). The correlations between VEGF-C expression and MVD, LVD, as well as other clinicopathological features were measured.

RESULTSAlthough no correlation between VEGF-C expression and tumor location, histological grade, or gender of the patients was observed (P > 0.05), OSCC patients with more advanced clinical stages and lymph node metastasis were prone to have high expression of VEGF-C (P = 0.015 and P < 0.001, respectively). Cases with high-expression of VEGF-C also showed significantly more often higher LVD (P = 0.001) but not MVD (P = 0.125). In addition, cases with lymph node involvement presented higher LVD than other cases (P = 0.026).

CONCLUSIONVEGF-C may promote lymph node metastasis by inducing lymphangiogenesis in OSCC.

Carcinoma, Squamous Cell ; Humans ; Lymph Nodes ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; Mouth Neoplasms ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor C

To understand the genetic characteristics of hemagglutinin (HA) and neuraminidase (NA) of type B influenza viruses in Guangzhou in 2006, three virus strains from etiology surveillance and seven strains from outbreaks were investigated. Genome RNAs of type B influenza viruses were extracted and reverse-transcripted into cDNAs using random primers. The whole-length DNA of HA and NA were amplified by polymerase chain reaction (PCR), cloned into T-A plasmid and sequenced, and analyzed phylogenetically by DNAstar software. The results showed that the HA of type B influenza viruses were similar and the homology were more than 99%. The type B influenza viruses belong to Victoria lineage. The NA of the type B influenza viruses were similar and the homology were more than 98%. Phylogenetic trees of HA and NA showed that the isolates from etiology surveillance formed a cluster, and the isolates from outbreaks were separated from the cluster. The homology of the type B influenza viruses with B/Shanghai/361/2002, which is the WHO recommended influenza vaccine strain in 2005-2006, were 88.9%-89.7%. It suggested the protective effect of influenza vaccine against type B influenza viruses used in 2005-2006 in Guangzhou may not be afforded.
China ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza B virus ; genetics ; Neuraminidase ; genetics ; Phylogeny ; Time Factors

Electrospray ionization is most commonly used in mass spectrometry for analysis of biological molecules such as peptides and proteins. However, peptides and proteins ions produced by electrospray ionization usually have multiple charges, and produce multiple spectrum peaks, making the mass spectrum complicated. Gas phase proton transfer ion/ion reaction can effectively regulate the charge states of peptides and proteins ions after electrospray ionization, simplifying the spectrogram, and thus is significant to the analysis of complex proteins and peptides samples. In this study, a dual-polarity linear ion trap ( LIT) mass spectrometer was used to control the charge state of peptide ions by proton transfer ion/ion reaction. The detection effect of the method was verified by using glutathione ( oxidation type) , oxytocin and dynorphin as typical samples. The results showed that this method could remove excess charges in the positive ions. When injecting enough negative ions for eaction, peptide ions with multiple charge valence state could be reduced to the minimum, therefore simplify the spectrogram effectively.

Objective To explore the therapeutic effects of endovenous laser ablation (EVLA) combined with percutaneous continuous circumsuture (PCCS) and EVLA in treating severe great saphena varicose. Methods A total of 60 patients with unilateral great saphenous varicose level C5-C6 were randomly divided into control group and experimental group according to the CEAP system. Control group was given EVLA surgery while experimental group was given EVLA+PCCS surgery. Data of operation time, hospital stay, intraoperative blood loss, the rate of ulcer healing, variceal recurrence rate and postoperative complication rate within 6 months after operation were compared between two groups. Results The mean operative time and intraoperative blood loss were lower in the experimental group than those in the control group ( P<0.05). There were no significant differences in hospital stay, ulcer healing rate and recurrence rate between two groups ( P>0.05). No deep venous thrombosis was found after treatment in two groups. The occurrence rates of skin burns and subcutaneous ecchymosis were significantly lower in the experimental group than those in the control group ( P<0.05). There were no significant differences in the incidence rates of other complications between two groups (P>0.05). Conclusion EVLA combined with PCCS in the treatment of severe saphenous varicose veins can significantly shorten the operation time, reduce the amount of bleeding, reduce the incidence rates of skin burns and subcutaneous ecchymosis on the premise of promising cure rate and recurrence rate. Overall, the combination therapy is superior than monotherapy.

Objective To study the clinical efficacy of bridge baton fixation with arthroscopic surgery in treatment of calcaneal fractures. Method From May 2013 to November 2014, 18 cases suffered from calcaneal fracture with arthroscopic surgical bridge baton fixation treatment in our hospital served as the object of the study. They were retrospectively analyzed during preoperative, postoperative the joint surface, the Bohler angle and Gissane angle , mean follow-up of 12 months, Maryland Foot Score were compared by the analysis. Results All cases were followed up for 12 months averagely. Joint surface flatness of 18 cases were significantly improved, Bohler angle, Gissane angle were also significant differences (P < 0.05). The excellent rate of Maryland score was 86.8%. Conclusion The bridge baton fixation with arthroscopic surgery for treatment of calcaneal fractures is certain effective and minimally invasive. Arthroscopy can restore the articular surface. The foot function recovery of the patients is satisfied.

OBJECTIVETo explore the intrinsic connection between activation of classical nuclear factor-κB (NF-κB) pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells.

METHODSHuman lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines.

RESULTSGefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines (P<0.05 or 0.01). Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells (P<0.05) and HCC827 cells (P<0.01). The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines (P<0.05). In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions (P<0.05 or 0.01), and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65 (P<0.01 or 0.01).

CONCLUSIONThe activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.

OBJECTIVE: To evaluate clinical effect of autologous osteochondral transplantation in treating localized knee cartilage defects. METHODS: Fifteen patients with knee cartilage defects were treated by autologous osteochondral transplantation from January 2007 to January 2008, including 8 females and 7 males, aged from 23 to 45 years old. Preoperative and postoperative KSS score at 10 years were compared. RESULTS: All patients were followed up for 10.0 to 10.7 years, with an average of(10.2±0.3) years. Clinical score of KSS was improved from 38.86±4.09 to 85.07±2.19 at 10 years after operation(<0.05), functional score increased from 3.33±4.88 to 82.67±4.58 at 10 years after operation(<0.05), KSS score was improved form 42.20±7.84 befor operation to 167.73±6.29 at 10 years after operation, and had statistical differences before and after operation. While there was no statistical difference in stability of knee joint(>0.05). All patients had no other complications. CONCLUSIONS Through long-term follow-up of patients with cartilage defect in knee treated by autologous bone cartilage transplantation showed that this method could effectively improve function of knee joint and alleviate pain. So it is an effective method for repair of osteochondral defect.
Adult ; Bone Transplantation ; Cartilage, Articular ; Female ; Follow-Up Studies ; Humans ; Knee Joint ; Male ; Middle Aged ; Osteochondritis Dissecans ; surgery ; Transplantation, Autologous ; Young Adult

Objective To explore the intracranial distribution of bone marrow-derived mesenchymal stem cells (BMSCs) and the ability of BMSCs shifting to glioma tissue.Methods We isolated BMSCs from the rats and constructed a BMSCsRL model that can stably express Renilla luciferase (RL).And 9L glioma cells marked with PKH26 were implanted into the brain parenchyma of Fischer rat using stereotactic surgery;7 d after that, the BMSCsRL was implanted into the contralateral brain parenchyma.The intracranial distribution of BMSCsRL was detected by using Xenogen bioluminescance imaging (BLI);at the same time,the migration of BMSCsRL into the glioma tissue was observed using Transwell plates.Results Phenotypical properties of the isolated BMSCs were CD90 and CD44 positive.BMSCs could be targeted to glioma tissue.In vivo BLI showed that the BMSCs shifted to the glioma tissue 0,7 and 14 d after transplantation and the junction area between tumor tissue and normal tissue was much more obvious than the other areas.Conclusion These results confirm the migratory capability of BMSCs over considerable distances, suggesting that BMSCs can be used as a delivery vehicle for targeted therapy of glioma.