Objective: To investigate the regulation of respiratory syncytial virus CTL epitope fused with G protein antigen fragment G1 to the specific immunoresponses. Methods: The recombinant plasmid pET-DsbA-G1 or pET-DsbA-G1F/M2 was transferred into E.coli BL21(DE3) and the fusion protein DsbA-G1F/M2 or DsbA-G1 was expressed.The expressing product was induced and purified by affinity chromatography. The two proteins were used to immunized BALB/c mice i.p, respectively. Serum and spleen cells were collected regularly. RSV-specific CTL responses were measured by MTT, IgG and IgG1 and IgG2a antibodies by ELISA, neutralizing antibodies by plaque reduction assay. Results: The recombinant proteins were expressed successfully and the purity was over 90% after purified by affinity chromatography. The protein DsbA-G1F/M2 induced significant RSV-specific CTL responses, while DsbA-G1 without CTL epitope did not induce detctable CTL responses. Strong IgG antibody responses were elicited,indicated by both. IgG1 and IgG2a titers induced by DsbA-G1F/M2, while only IgG1 was induced by DsbA-G1.The ratio of IgG1/IgG2a was downregulated significantly. Both antigens induced high level of neutralizing antibodies, but the latter was a little lower. Conclusion: DsbA-G1F/M2, the fusion protein of a CTL epitope and G protein fragment G1 can induce both cellular immunity and humoral immunity. The activation of CTLs downregulates the ratio of IgG1/IgG2a.The more balanced immunoresponse is advantageous for improving safety of the candidate vaccine.

[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.

A reasonable and practicable quality standard was developed for mori liquid extract from different sources by TLC, HPLC and fingerprint technology. In TLC method, the compounds were separated on polyamide film using glacial acetic acid-water (1: 3) as mobile phase at a UV wavelength of 365 nm. All qualified samples had the spots of the same color as the control herb and substance. The RP-HPLC method was used to determine the content of mulberroside A with mobile phase of methanol-water (25: 75) at a wave-length of 326 nm. The mulberroside A was in good linear with a regression equation of Y = 46.965X (r = 0.999 6) in the range of 4.6 - 228 mg x L(-1). In 14 batches of samples, the mulberroside A in 4 batches of them was less than 0.5 g x L(-1), and was more than 2.0 g x L(-1) in the other batches. It was suggested that the content limit of mulberroside A should be no less than 1.5 g x L(-1). The HPLC fingerprints were evaluated by the similarities. It has found that the similarities of different mori liquid extracts were very low and the chemical diversity of mori cortex was the major factor of similarity. Moreover, the process impact was minimal. Thus the fingerprint was not included in this quality standard.
China ; Chromatography, High Pressure Liquid ; Disaccharides ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Morus ; chemistry ; Quality Control ; Stilbenes ; chemistry ; isolation & purification

BACKGROUND: Mesenchymal stem cell (MSC) transplantation has been gradually developed to improve the prognosis of cerebral infarction and its sequelae in clinical trials, which has been identified as effective and safe. A small sample size, however, results in the lack of evidence-based medical evidence.OBJECTIVE: To systematically review the efficacy of MSC transplantation on the prognosis of cerebral infarction. METHODS: In order to collect randomized controlled trials (RCTs) of MSC transplantation for the prognosis of cerebral infarction, we searched Cochrane Library, PubMed, Ovid, CBM, CNKI, WanFang, and VIP Data from its inception to November 2016. Articles addressing MSCs transplantation alone or with conventional drug treatment and/or rehabilitation training versus conventional drug treatment alone or with rehabilitation training were included. Two authors independently screened the literature according to the inclusion and exclusion criteria, extracted data, and assessed the risk of bias. Thereafter, qualitative description and Meta-analysis were performed. RESLUTS AND CONCLUSION: Ten RCTs involving 626 cerebral infarction patients were included in the Meta-analysis. The results showed that the MSCs group was superior to the control group with statistical significance in the daily life ability(Barthel index)[MD=20.06, 95%CI(9.95,30.18),P=0.000 1],motor function(Fugl-Meyer scale)[MD=14.60,95% CI(12.96,16.25),P<0.000 01],personal disability (functional independent measure)[MD=15.16,95%CI(9.06,21.26),P<0.000 01]and neurological deficit score(National Institute of Health stroke scale)[MD=-2.59,95% CI(-3.14,-2.05),P<0.000 01].Low fever and mild headache were reported by four included studies,and waist pain was only by one study, but these symptoms went away by themselves or after symptomatic treatment. Subgroup analysis suggested that MSCs from the bone marrow were superior to those from the umbilical cord and cord blood, but showed a greater heterogeneity. It is suggested that the MSC transplantation ameliorate the prognosis in patients with cerebral infarction, significantly improve the activities of daily living, motor function, personal disability and neurological function, with no presence of serious adverse effects. However, high-quality studies with large sample size are required for further investigation on the clinical application of MSC transplantation.

Undifferentiated and differentiated 3T3-L1 adipocytes were treated with 100 ng/ml tumor necrosis factor-?(TNF-?),and peroxisome proliferator-activated receptor-?2 (PPAR-?2) mRNA expression and adiponectin secretion in cultured cells were measured.The results showed that TNF-?suppressed PPAR-?2 mRNA expression and adiponeetin secretion in 3T3-L1 adipocytes (P

Objective To explore the relationship between wild rodent plague and human in wild rodent plague foci of the northwestern area in Yunnan to probe the possible transmission mechanism of wild rodent plague to human. Methods Data of component ratio of rodents and fleas was collected in different areas from 1985 - 1995. Activities and habits of residents regarding the way they keep cats and dogs and parasitic fleas and free fleas indoor were investigated, the dog serum was collected for detecting F1 antibody. Results Eothenomys miletus were main rodents in farmland and shrub, accounting for 48.00% (4753/9902) and 54.50% (4282/7857), Apodemus chevrieri were main rodents in garden, being 50.47% (1332/2639). The component ratio of Neopsylla specialis specialis was 13.31%(229/1720), 12.31%(1678/13 739) and 10.87%(957/8802) respectively in garden, farmland and shrub, higher than in indoor. The component ratio of Frantcpsylla spodix was 39.88% (686/1720), the highest in garden. Thirty-two per cent (32/100) of residents kept cats,in which 63% (20/32) with cat fleas, 68% (68/100) of villages kept dogs, in which 76%(52/68) with fleas. Eighteen parasitic fleas were caught from 43 dogs with a flea index of 0.119 and a rate for fleas of 11.63%, 7 pulex were collected from 17 indoor. Forty-three blood serum samples were obtained from dogs, among which 3 were positive blood serum. Conclusions Residents touch affected animals or media in different situations. The possibility of transmission for wild rodent plague to human exists in loci in a chain of wild rodent plague → fleas or predation → homebred animal plague (cats or dogs) →touching or respiratory → human.

OBJECTIVETo investigate the antagonistic effects of three species of oral Streptococcus on the growth of oral Saccharomyces albicans in vitro.

METHODSDirect inoculation method, reverse inoculation method and mixed culture methods were respectively chosen to observe the changes of Saccharomyces albicans colony formation on the effects of Streptococcus mutans, Streptococcus sanguis and Streptococcus salivarius.

RESULTS1) No clear inhibition zone was observed in each of the groups by direct inoculation method. 2) Compared with the control groups, Saccharomyces albicans colony formation on soft agar of Streptococcus sanguis decreased significantly (P < 0.05). 3) Mixed culture method results showed that Streptococcus mutans could inhibit the growth of Saccharomyces albicans significantly at different time points (P = 0.001). 4) Under the action of bacteria culture supernatant, the count of Saccharomyces albicans in experiment groups showed statistical significance when compared with the control groups at 24, 48, 72 h (P = 0.001); The differences among the experimental groups were of no statistical significance at majority times (P > 0.05).

CONCLUSIONStreptococcus mutans, Streptococcus sanguis, and Streptococcus salivarius could obviously inhibit the growth of Saccharomyces albicans in vitro. However, it is still unclear that among which the inhibition effects is stronger. The antagonistic effects is weakened gradually.

In Vitro Techniques ; Saccharomyces ; Streptococcus ; Streptococcus mutans ; Streptococcus sanguis

OBJECTIVEThe aim of this study was to investigate whether the three species of oral Actinomyces have inhibitory effects on the growth of oral Candida albicans in vitro.

METHODSStraight o'clock method was used to observe the bacteriostasis circle. Reverse o'clock and mixed culture method were used to study the quantitative changes of Candida albicans colony respectively.

RESULTS(1) None of the groups had been viewed the bacteriostasis circle. (2) Compared with control groups, there was a significant decrease of Candida albicans colony on Actinomyces viscosus TPY soft agar (P < 0.05). Actinomyces naeslundii and Actinomyces odontolyticus TPY soft agar were both devoid of obvious Candida albicans colony (P < 0.01). The former group (Actinomyces viscosus) and the two latter groups (Actinomyces naeslundii and Actinomyces odontolyticus) showed a striking contrast (P < 0.01). (3) Compared with control groups, a decrease of Candida albicans showed up in the mixed culture, and the difference was significant (P < 0.05). The discrepancies among the three experimental groups were of no statistical value (P > 0.05).

CONCLUSIONOral Actinomyces viscosus, Actinomyces naeslundii and Actinomyces odontolyticus could inhibit the growth of Candida albicans in vitro. However, which of them contributed more to the inhibitory effects was still not affirmed.

Actinomyces ; Actinomyces viscosus ; Candida albicans ; In Vitro Techniques

OBJECTIVETo investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1alpha level of HDPC supernatants stimulated by lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-alpha), and to explore the role of SDF-1alpha on the proliferation and the migration of HDPC.

METHODSThe expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique. The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-alpha of different concentrations for 48h and then the SDF-1alpha level was assayed by quantitative sandwich ELISA. Meanwhile, the effects of recombinant human SDF-1alpha (rhSDF-1alpha) on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.

RESULTSCXCR4 was expressed in cytomembrane of HDPC and SDF-1alpha was secreted into their normal cell supernatants with a concentration of (4513.55 +/- 962.92) ng/L. The secretion of SDF-1alpha was both significantly decreased by stimulation with LPS and TNF-alpha (P < 0.05). In addition, rhSDF-1alpha stimulated the HDPC proliferation at the concentrations of 50, 100, 200 microg/L (P < 0.01) and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50, 100 microg/L (P < 0.05).

CONCLUSIONSSDF-1alpha accelerated the proliferation and the migration of HDPC which expressed CXCR4. SDF-1-CXCR4 axis may play a role in repair of pulp injury.

Cell Movement ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12 ; metabolism ; Dental Pulp ; cytology ; metabolism ; Humans ; Receptors, CXCR4 ; metabolism

OBJECTIVETo investigate the expression of macrophage migration-inhibitory factors (MIF) in clinically healthy and inflamed human pulp tissues and the effects of rhMIF on the proliferation of human dental pulp cells (HDPC).

METHODSImmunohistochemistry was used to detect the localization of MIF expression in clinically healthy pulp and inflamed pulp tissues. Quantitative real-time polymerase chain reaction (PCR) was performed to evaluate the mRNA levels of MIF in pulp specimens. In addition, the culture supernatants of HDPC were collected after HDPC was stimulated by lipopolysaccharide (LPS) for 24 h, and then the MIF levels were assayed by quantitative sandwich enzyme-linked immunosorbent assay. Meanwhile, the effects of rhMIF on the proliferation of HDPC at different concentrations for 24 and 48 h were observed by cell counting kit-8 (CCK-8).

RESULTSMIF was mainly distributed in odontoblasts of healthy pulp tissue, however, in inflamed pulp tissue, it was widely detected in fibroblasts, inflammatory infiltrates and endothelial cells as well as odontoblasts. Quantitative real-time PCR showed that there was no significant difference in MIF mRNA levels between inflamed pulps and healthy pulps (P > 0.05). Additionally, the secretion of MIF was significantly increased by stimulation with LPS at the concentration of 0.1 and 1.0 mg/L [(1772.58 ± 495.05), (1692.58 ± 337.45) ng/L] (P < 0.05), and the concentration was (1048.53 ± 161.81) ng/L in control group. rhMIF stimulated the HDPC's proliferation at the concentration of 10, 30, 60 µg/L for 24 and 48 h.

CONCLUSIONSMIF was expressed in pulp tissue and its expression was increased after stimulation by LPS. rhMIF increased the proliferation of HDPC. These results suggest that MIF may be involved in the process of pulpal inflammation.

Cell Proliferation ; Cells, Cultured ; Dental Pulp ; cytology ; pathology ; Dose-Response Relationship, Drug ; Humans ; Lipopolysaccharides ; administration & dosage ; pharmacology ; Macrophage Migration-Inhibitory Factors ; genetics ; metabolism ; Pulpitis ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction