Objective To investigate the effect of cold atmospheric plasma (CAP) on the healing of skin ulcers using Balb/c mice. Methods Wounds with a diameter 6 mm were created on each side of the backs of BalB/c mice ( n = 150) using a punch bioptome. The mice were assigned randomly into a control group ( wounds healed naturally), a laser group (wounds treated with a He-Ne laser for 10 min daily) and a CAP group (wounds treated with CAP for 10 min daily). Wound healing was evaluated on postoperative days (PODs) 4, 7, 10 and 14 in terms of percent wound closure. Ten mice per group were sacrificed on each of the evaluation days. Both wounds were removed and a histological examination was conducted. A scoring system was used to evaluate the wounds. The expression of vascular endothelial growth factors (VEGFs) in the wounded tissue was detected by using immunohistochemical methods on POD 7. The results were quantified using an HPIAS-1000 system. Results Compared with the control group, the average percentage of wound healing was significantly greater in the CAP group on PODs 7 and 10. The average scores on the histological examination were significantly higher in the CAP group on PODs 7, 10 and 14. Compared with the other two groups, the expression of VEGF was up-regulated significantly in the CAP group.Conclusions CAP can positively affect the wound healing process. This might be related to the up-regulation of VEGF in the wounded tissues.

Objective To explore the related factors influencing the prognosis of children with Tourette syndrome ( TS) . Methods We collected 420 outpatient cases of TS children in the department of pediatrics of Guangdong Provincial Hospital of Traditional Chinese Medicine from January of 2007 to October of 2010 as the research objects. Using the unified survey questionnaire, we observed the influencing factors of TS prognosis such as sex, onset age, mother’s pregnancy situation, the perinatal period, partiality for food intake, family relationship, the first symptom, the severity of disease, underlying diseases, comorbidities ( such as attention deficit hyperactivity disorder, anxiety disorder, and obsessive-compulsive disorder) , family history of psychiatric or neurological diseases, trace elements levels, electroencephalogram (EEG), antistreptolycin O (ASO), course of disease, and maintenance treatment. The related factors of prognosis was analyzed with single factor and multiple factors Logistic regression analysis. Results Among the 420 cases of TS children, 396 cases were included into the final analysis, 24 were lost and the follow-up lost rate was 5.7%. The remission rate of TS was 78.3%, and the uncured rate was 21.7%. The results of preliminary screening of the influencing factors by single factor Logistic analysis showed that the related influencing factors for TS prognosis were 12, and they were course of diseases, abnormal birth history, father’s education level, mother’s education level, upbringing methods, family history of psychiatric or neurological diseases, underlying disease history, comorbidities history, abnormal ASO, the severity of disease, the frequency of disease relapse, and the medication history of western medicine (P<0.05) . And then the obtained 12 factors were analyzed by the multiple fac tors Lo gistic regression analysis, the results showed that upbringing methods, comorbidities history, the severity of disease, and the frequency of disease relapse were correlated with TS prognosis ( P < 0.05) . Conclusion TS children will have poor prognosis when their parents spoil, indiscipline, beat and scold, and dictate them, or when the children have severe illness state, frequent recurrence of the disease, and the history of comorbidities ( atten tion deficit hyperactivity disorder, anxiety disorder, obsessive-compulsive disorder, etc.) .

Objective To study the expression of cellular FLICE inhibitory proteins(c-FLIP)in peripheral blood B lymphocytes in patients with systemic lupus erythematosus (SLE)and its correlation with clinical features.Methods Blood samples were obtained from 53 patients with SLE and 30 normal human controls.Flow cytometry and ELISA were performed to measure the expression of c-FLIP in pefipheral blood B lymphocytes and serum levels of IL-4 and IL-10,respectively.Relevant laboratory examinations were carried out for these patients.SLE disease activity index(SLEDAI)score was calculated for patients.Results The positivity rate of c-FLIP in B lymphocytes was 3.11%±0.70%in 18 patients with active SLE.significantly higher than that in 35 patients with inactive SLE (0.78%±0.28%)and normaI controls(0.68%±0.12%),while no statistical difierence was found between inactive patients and controls(t=1.56,P＞0.05).In SLE patients,the expression of c-FLIP showed a positive correlation with SLEDAl score(r=0.96.P＜0.05),erythrocyte sedimentation rate(r=0.96,P＜0.01),serum level of C reactive protein(r=0.92.P＜0.01)and the titer of antinuclear antibodies(r=0.86,P＜0.01),whereas in 36 patients with leucopenia.a negative correlation was noticed between white blood cell count and the expression level of c-FLIP(r=-0.94,P＜0.0 1).The 23 patients with lupus nephritis had a higher level of c-FLIP than those without lupus nephritis(3.04%±1.09%vs 1.76%±1.09%,t=4.23,P＜0.05).Additionally.the expression of c-FLIP positively correlated with the serum level of IL-4 and IL-10(r=0.80,0.89.respectively,both P＜0.01).Conclusions In patients with active SLE,the expression of c-FLIP is upregulated in peripheral blood B lymphocytes,and positively correlated with the severity of SLE as well as the serum level of IL-4 and IL-10.The upregulation of c-FLIP in B cells might play a certain role in deficient apoptosis or clearance of activated B cells in SLE.

Objective To investigate the effect of recombinant murine interleukin-23(rIL-23)on systemic candidiasis in a murine model.Methods A cyclophosphamide-induced immunosuppressed murine model of systemic candidiasis was established.The mice were divided into control group and rIL-23 treatment group.Colony forming units(CFU)of the kidney and spleen were determined by using plating dilution method.The histopathological changes and degree of infection of the kidney and spleen were graded.Meanwhile,the levels of interferon gamma(IFN-γ)in the spleen were measured by enzyme-linked immunosorbent assay.Results On the 2nd,3rd and 7th day after Candida albicans infection the number of CFU of the fungi in the kidney in the control group was significantly greater than that in rIL-23 treatment group(P＜0.01).The number of CFU of the fungi on the 2nd,3rd and 7th day after Candida albicans infection in the spleen in control group was also greater than that in rIL-23 treatment group,but without statistically significant difference(P＞0.05).The scores of histopathological changes in the kidney in rIL-23 treatment group were lower than those in control group(P＜0.01),and the degree of infection was milder in rIL-23 treatment group.The scores of histopathological changes in the spleen in rIL-23 treatment group were also lower than those in control group,but without statistically significant difference(P＞0.05).The levels of IFN-γ in the spleen on the 2nd,3rd and 7th day after infection in rIL-23 treatment group were significantly higher than those in control group(P＜0.01).Conclusion rIL-23 has protective effect on murine systemic candidiasis.

Objective To observe the clinical efficacy of Q-switched Alexandrite laser at 752 nm in the treatment of nevus of Ota. Methods A total of 1985 cases of nevus of Ota were treated with the Q-switched Alexandrite laser PhotoGenica HT10, and then the ages, frequency of treatment and interval of treatment were analyzed. Results The excellent effective rate was 97.88 %, and the total effective rate was 100 % in 1985 cases. Most patients in all age group received the excellent effects, however, there was no significant difference between the groups. Most patients acheived the excellent effect after 4 to 5 treatments, and very few patients (0.8 %) needed over 10 treatments; the rate ofpatient who needed 1-3 treatments or 6-10 treatments was 18. 2 % and 25.8 %, respectively. The patients had the most excellent efficacy in the group that the interval of two treatments was 4 to 6months, however, there was no significant difference between the group of the interval of two treatments over 6 months. In our study, there were only a few cases (4.48 %) with slight side reaction,such as temporary pigmentation and hypopigmentation and scar. Conclusions 752 nm Q-switched Alexandrite laser is one of effective and safe treatments for nevus of Ota.

In this study, the sterilizing effect of atmospheric pressure nonequilibrium plasmas (APNPs) on Neisseria gonorrhoeae (N. gonorrhoeae) was preliminarily examined and the possible mechanisms were explored. N. gonorrhoeae FA1090, FA19 and MS11 were treated by APNPs and their survival rate was analyzed by using CFUs counting and structurally studied by laser scanning confocal microscopy. The morphological changes of bacterial cell membrane and wall were studied under TEM. Our results showed that APNPs had strong sterilizing effect on N. gonorrhoeae. The survival rate of MS11 in N. gonorrhoeae liquid medium was 60.65% after disinfection with the APNPs for 5 min, whereas, the survival rate of FA19 was 92.60% and the rate of FA1090 was 96.40%. The survival rate of MS11 was 21.13% after exposure to APNPs for 6 min, whereas the survival rate of FA19 was 31.60% and the rate of FA1090 was 91.00%. N. gonorrhoeae was structurally damaged after treatment with APNPs. It is concluded that APNPs is able to effectively and quickly kill the N. gonorrhoeae, and the killing effect is related to the architectural damage of cell membrane.

To identify the genomic species of Neisseria gonorrhoeae, evaluate the difference between two molecular epidemiological methods and examine the relationship between sex partners and genotypes of bacteria, 24 strains of Neisseria gonorrhoeae isolated from the outpatients with gonorrhea were identified by using the Opa genotyping and NG-MAST genotyping and the relationship between genotypes and phenotypes was studied. Twenty-four strains of Neisseria gonorrhoeae fell into 10 ST genotypes by NG-MAST genotyping, whereas these strains were classified into 12 OT Opa genotypes by Opa genotyping. A new epidemic strain of ST genotype (217-86% homologisation 178) in China was identified. It is concluded that genotypes of each pair of strains from a pair of patient/ sex partner besides 45/46 are the same, indicating that contagious infection take place between patient and the sex partner. Opa genotyping was more effective than NG-MAST genotyping in identifying the genomic species of Neisseria gonorrhoeae. ST genotype could be further classified into different Opa-types.

TNFR-Fc is an important fusion protein that has great potential in therapeutic and diagnostic applications. We developed an efficient fed-batch process for GS-CHO cells to produce TNFR-Fc. The rationale of this fed-batch process relies on the supply of sufficient nutrients to meet the requirements of cell metabolism. The optimal feed medium was designed through ration design. A metabolically responsive feeding strategy was designed and dynamically adjusted based on the residual glucose concentration determined off-line. In this process, the maximal viable cell density and antibody concentration reached above 9.4x10(6) cells/mL and 207 mg/L, respectively. Compared with the batch process, the newly developed fed-batch process increased the cell yield by 3.4 fold and the final antibody concentration by 3 fold. This fed-batch process would therefore facilitate the production of therapeutic antibody by GS-CHO cells.
Animals ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media ; Etanercept ; Glucose ; analysis ; Immunoglobulin G ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics

The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice. Two 6-mm wounds along the both sides of the spine were created on the back of each mouse (n=80) by using a punch biopsy. The mice were assigned randomly into two groups, with 40 animals in each group: a non-thermal plasma group in which the mice were treated with the non-thermal plasma; a control group in which the mice were left to heal naturally. Wound healing was evaluated on postoperative days (POD) 4, 7, 10 and 14 (n=5 per group in each POD) by percentage of wound closure. The mice was euthanized on POD 1, 4, 7, 10, 14, 21, 28 and 35 (n=1 in each POD). The wounds were removed, routinely fixed, paraffin-embedded, sectioned and HE-stained. A modified scoring system was used to evaluate the wounds. The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group, earlier than in control group in which acute inflammation reached a peak on POD 7, and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7 (P<0.05). The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group (P<0.05). The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10 (P<0.05 for all). The count of bacterial colonies was 10(3) CFU/mL on POD 4 and <20 CFU/mL on POD 7, significantly lower than that in control group (10(9) CFU/mL on POD 4 and >10(12) CFU/mL on the POD 7) (P<0.05). It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colonization.

In this study,the sterilizing effect of atmospheric pressure nonequilibrium plasmas (APNPs)on Neisseria gonorrhoeae(N.gonorrhoeae)was preliminarily examined and the possible mechanisms were explored.N.gonorrhoeae FAI090,FA19 and MS11 were treated by APNPs and their survival rate was analyzed by using CFUs counting and structurally studied by laser scanning confocal microscopy.The morphological changes of bacterial cell membrane and wall were studied under TEM.Our results showed that APNPs had strong sterilizing effect on N.gonorrhoeae.The survival rate of MS11 in N.gonorrhoeae liquid medium was 60.65% after disinfection with the APNPs for 5 min,whereas,the survival rate of FA19 was 92.60% and the rate of FA1090 was 96.40%.The survival rate of MS11 was 21.13% after exposure to APNPs for 6 min,whereas the survival rate of FA19 was 31.60% and the rate of FA1090 was 91.00%.N.gonorrhoeae was structurally damaged after treatment with APNPs.It is concluded that APNPs is able to effectively and quickly kill the N.gonorrhoeae,and the killing effect is related to the architectural damage of cell membrane.