Objective To study the antibacterial effect of borneol, honey, gentamicin in Ruchuang Bingmi hydropathic compress agent.Method Staphylococcus aureus(S.aureus), Escherichia coli(E.coli), Pseudomonas aeruginosa(P.aeruginosa) were used to produce the third phase of infection decubitus animal models, respectively.Vaseline as the control group, divided into 12 groups:Vaseline-S.aureus, Vaseline-E.coli, Vaseline-P.aeruginosa;Honey-S.aureus,Honey-E.coli,Honey-P.aeruginosa;Borneol -S.aureus;Borneol -E.coli;Borneol-P.aeruginosa;Gentamicin-S.aureus;Gentamicin-E.coli;Gentamicin-P.aeruginosa;6 rabbits in each group.Honey, borneol, gentamicin was made into a gauze in treatment for decubitus .Organizations do strain identification and colony counts was observed before and after taking the treatment.ResuIts Borneol group ( F =11.059,P<0.01).,there was differences of each groups count cultured by borneol;Time ×Strains(F=11.281,P=0.009),there was no significant interaction between the two groups;gentamicin(F=7.99,P=0.000),gentamicin culture showed a difference in each group count;Time ×Strains(F=12.531, P<0.07),interaction between the two groups showed significant.Borneol has no antibacterial effects on P.aeruginosa , had a certain antibacterial activityon S.aureus and E coli;gentamicin had good antibacterial effect on the three kinds of bacteria, and against P.aeruginosa was particularly significant.ConcIusion The antibacterial activity of gentamicin is better than single herbs borneol in Ruchuang Bingmi hydropathic compress agent, honey has no antibacterial effect.

Background Deficiency of neurotrophic factor is associated with the damage of optic nerve in glaucoma.Reaserches showed that ectopically applied neurotrophic factor has a transient neuroprotective effect in glaucoma model,and the viral expression of adeno-associated neurotrophie factor may provide long-term supplementation of neurotrophic factor and neuroprotection in tissues.Objective The present study Was to investigate the neuroproteetive effect of adeno-associated viral(AAV)-mediated brain-derived neurotrophic factor (BDNF)on retinal ganglion cells(RGCs)in DBA/2J mice with experimental glaucoma. Methods 10 clean DBA/2J mice were administered intravitreal injection with 1 microliter of AAV-BDNF-GFP in the left eyes at the age of 6 months,and the right eyes were injected with the same volume of saline solution as control.Intraocular pressure (IOP)was measured with Tonolab in the mice every month.Retinas were obtained after 3 months for the investigation of GFP expression in RGCs using fluorescence microscopy.Immunohistochemistry Was performed by applying TUJ1 and Cy3 antibodies to identify surviving RGCs. Results The IOP of DBA/2J mice were 11.90 mmHg and 11.40 mmHg in the right eyes and left eyes,respectively,at 4 months.The IOP of DBA/2J mice began to rise at 5 months and reached its peak in 8 month-old mice.There was no statistically significant difference in IOPs between the right eyes and the left eyes from 4 month-through 9 month-old mice(t=-1.78-0.61,P=0.11-0.90).Three months after intrlavitreal injection of AAV-BDNF-GFP,GFP was positively expressed in RGCs of retinas with the expression rate of 46.33%±8.08%.The over-expression of BDNF led to more RGCs survival than the control eyes (3168.13±1319.33/mm2 vs 2024.81±796.38/mm2,t=2.75,P=0.02). Conclusion These data suggest that BDNF can exert a protective effect in DBA/2J glaucoma mice.

OBJECTIVETo explore a new method for early avascular necrosis of femoral head (AVNFH) therapy.

METHODSSixty-nine AVNFH New Zealand adult rabbits were randomly divided into three groups with equal number. In Group A, deproteinized bone (DPB) that absorbed with recombinant plasmid pcDNA3.1-hVEGF165 was implanted into the drilled tunnel of necrotic femoral head. In Group B, only DPB was implanted. In Group C, only tunnel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained at postoperative 1, 2, 4, 8, 16 weeks. The expression of VEGF165 and collagen I was detected by immunohistochemistry. Bone formation was detected generally by X-ray. Angiogenesis and the repair of the femoral head were observed histologically.

RESULTSThe expression of VEGF 165 could be detected 2 weeks after implantation in Group A, but it was not observed in other groups. The result of collagen I expression had a significantly difference 2, 4 and 8 weeks after operation in Group A from those in other groups (P < 0.01). X-ray results indicated that there was more bone formation in Group A than in other groups. The regenerated capillary vessels staining result of necrotic femoral head in Group A was significantly different from those in other groups at postoperative 2 and 4 weeks (P < 0.01).

CONCLUSIONSTransfection of hVEGF165 gene enhances local angiogenesis and DPB-VEGF compound improves the repair of necrotic femoral head. Deproteinized bone grafting with VEGF gene transfer provides a potential method for the treatment of osteonecrosis.

Animals ; Bone Transplantation ; Collagen Type I ; analysis ; Femur Head Necrosis ; pathology ; physiopathology ; therapy ; Genetic Therapy ; Immunohistochemistry ; Neovascularization, Physiologic ; Rabbits ; Transfection ; Vascular Endothelial Growth Factor A ; analysis ; genetics

OBJECTIVETo study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution.

METHODSOverlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed.

RESULTSThe genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I.

CONCLUSIONThe nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.

Animals ; Cell Line ; China ; Cricetinae ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny

AIMTo establish methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein (rhTNFR-Fc).

METHODSBiological potency of rhTNFR-Fc was determined by neutralizing the bioactivity of TNF-alpha. rhTNFR-Fc samples were reduced by beta-mercaptoethanol and the peptide map was performed by tryptic digestion. Residual protein A and the host cell protein content were detected by ELISA. Anti-TNFR and anti-Fc antibodies were used in ELISA for detection of the rhTNFR-Fc content.

RESULTSThe quality control methods, such as bioassay, peptide map, residual protein A detection, were established and used for quality control of rhTNFR-Fc. The unit of rhTNFR-Fc (AU) was defined according to the international unit of TNF-alpha. The specific activity was up to 8 x 10(4) AU.mg-1. The requirements for quality control of rhTNFR-Fc were established.

CONCLUSIONThe methods and requirement were used for quality control of rhTNFR-Fc products.

Animals ; Biotechnology ; methods ; Cell Division ; drug effects ; Etanercept ; Immunoglobulin G ; biosynthesis ; chemistry ; pharmacology ; Mice ; Peptide Mapping ; Quality Control ; Receptors, Tumor Necrosis Factor ; biosynthesis ; chemistry ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; pharmacology ; Technology, Pharmaceutical ; standards ; Tumor Cells, Cultured

To establish methods and requirements for quality control of rhLFA3-IgG1, biological potency of rhLFA3-IgG1 was determined by CD2 molecule competitive binding assay on Jurkat cell surface. Purity of rhLFA3-IgG1 was analyzed by SEC-HPLC and IEC-HPLC. Peptide mapping was preformed by tryptic digestion and RP-HPLC after sample reduced and carboxymethylation by DTT and indoacetic acid, respectively. CHO host cell protein and Protein A residual were detected by ELISA separately. The quality control methods and requirements, such as biological potency, the physical-chemical characteristic of rhLFA3-IgG1 had been established. The methods and requirements for quality control of rhLFA3-IgG1 showed advantages of assuring the products safety and efficacy, which can be used for routine quality control of rhLFA3-IgG1.
Binding, Competitive ; Biotechnology ; methods ; CD2 Antigens ; metabolism ; CD58 Antigens ; biosynthesis ; chemistry ; Chromatography, High Pressure Liquid ; Humans ; Immunoglobulin G ; biosynthesis ; chemistry ; Jurkat Cells ; Molecular Weight ; Peptide Mapping ; Quality Control ; Recombinant Fusion Proteins ; biosynthesis ; chemistry

OBJECTIVETo explore the follow-up visit, outcome and auxiliary diagnosis method on the cases with indeterminate antibody level measured by Western blotting as well as the related biological factors.

METHODSThe cases with indeterminate result were followed up according to the National Guideline for Detection of HIV/AIDS (2009) and samples were collected for HIV antibody detection, p24 antigen and nucleic acid were detected as a supplementary diagnosis at the same time. The samples were also be detected for HBV, HCV, TP, HTLV-I/II, ANA, and AFP, and the results were compared to that of screened positive and confirmed negative cases.

RESULTSA total of 73 were followed up successfully and taken a second HIV test, 25 cases were tested positive and 48 were tested negative for HIV during the follow-up period. For the 25 HIV positive cases, the HIV seroconversion rate was 100.00% at any time point when the interval between the first and returning detection was longer than 1 week. The major Western blotting bands for the cases with indeterminate result were p24 and gp160 and it was different between HIV positive and negative cases in Western blotting band profiles. The consistency and sensitivity of nucleic acid detection were higher than 90.00%, and were higher than that of p24 antigen (69.09% (38/55) and 27.27% (6/22)) (χ(2)(consistency) = 6.875, χ(2)(sensitivity) = 18.893, P < 0.05). The positive rates of ANA and AFP of indeterminate cases excluded from HIV infection were 20.83% (10/28) and 6.25% (3/48) and higher than that of screened positive and confirmed negative cases (0.00%), the difference had statistic significance (χ(2)(ANA) = 19.430, χ(2)(AFP) = 5.520, P < 0.05).

CONCLUSIONIt is critical to get timely diagnosis for the indeterminate cases according to the new national guideline for detection of HIV/AIDS. Nucleic acid detection has higher application value as auxiliary diagnosis for HIV infection than p24 antigen. The increased levels of ANA and AFP may be the factors resulting in the nonspecific indeterminate results.

Antibodies, Antinuclear ; blood ; Female ; Follow-Up Studies ; HIV Antibodies ; blood ; HIV Infections ; diagnosis ; immunology ; Humans ; Male ; alpha-Fetoproteins ; analysis

Objective:To investigate the effects of mycophenolate mofetil ( MMF) on the differentiation and connective tissue growth factor(CTGF)and fibronectin(FN)expression of lung fibroblasts(LF)through interfering the transdifferentiation of LF into MF in vitro.Methods:LF of neonatal rat were cultured in vitro ,induced into MF by transforming growth factor-β1(TGF-β1),and treated with different concentrations of MMF ,which was 0μmol/L(control group),0.1μmol/L(low dose group),1μmol/L(middle dose group)or 10 μmol/L( high dose group ) .Morphology of LF and MF were observed by inverted phase contrast microscope , the expressions of vimentin and α-smooth muscle actin (α-SMA) were identified by immunofluorescence staining ,and then analyzed the effect of MMF on the transdifferentiation of fibroblasts .ELISA was used to detect the levels of connective tissue growth factor ( CTGF ) and fibronectin ( FN) .Results: LF was induced into MF by TGF-β196 hours later.The immune fluorescence performance of α-SMA in the lung fibroblasts revealed MMF could suppress the expression of α-SMA,but had no effect on the phenotype of myofibroblasts .The results of ELISA showed that the levels of CTGF and FN were significantly decreased compar with that of control group and was concentration -de-pendent ( P<0.05 ) .Conclusion: MMF can prevent lung fibroblasts from transdifferentiating into myofibroblasts and inhibit the expressions of CTGF and FN ,suggesting that MMF has anti-fibrosis effect and one of the mechanisms is by suppressing the expressions of CTGF and FN.

Objective To introduce and conduct effectiveness evaluation of peer-group level care and management mode in people living with HIV and AIDS(PLHAs)in Jing'an District, Shanghai. Methods We recruited PLHAs who were managed by Jing'an District and participated in the peer-group care activities from 2015 to 2016, and collected their data of general conditions, behavior, antiretroviral therapy, life quality, social support, mental status and medical modes, and then conducted effectiveness evaluation of the mode by multivariate logistic regression model and linear regression model. Results A total of 300 PLHAs were included with 150 cases in each group. PLHAs in target group had higher proportion of antiretroviral therapy[82.7%(124/150)and 64.7%(97/150)]. In addition, they got higher scores in mental health(48.5±9.2 and 43.9±10.6)and social support(28.7±8.2 and 24.8±6.6), and lower scores in depressive severity index(0.5±0.1)and avoidance of medical modes(16.1±3.0 and 17.0±2.5). Multivariate analysis showed that PLHAs in target group had less sex partners(zero partner, OR = 2.1, 95%CI:1.1-3.9;one partner, OR = 3.6, 95%CI:1.9-6.8)and increased condom use(no sex, OR = 5.3, 95%CI:2.4-11.7;complete use, OR = 10.6, 95%CI:4.7-24.1;random miss, OR = 5.1, 95%CI:1.8-14.8)in the last 12 months. Conclusion The effect of peer-group level care and management mode is significantly good in promoting behavior change, enhancing health concern and increasing antiretroviral therapy.

BACKGROUND: There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment. METHODS: We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes. RESULTS: Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100). CONCLUSION 5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.
Animals ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12/genetics* ; Fibroblasts ; Fluorouracil/therapeutic use* ; Humans ; Mice ; RNA, Messenger ; Receptors, CXCR4