Objective:To observe the effects of different doses of advanced glycosylation end products ( AGEs ) on bFGF expression of cultured rabbit M üller cells in vitro.Methods:Immunocytochemistry and transmission electron microscopy methods were used identified cultured M üller cell.Immunocytochemistry method was used to semi-quantitate bFGF expression of retinal Müller cells at 640 μl/2 000 μl AGEs conditions.We observed effects of AGEs and PKC inhibitor Calphstion C on bFGF mRNA expression .Results:640 μl/2 000 μl AGEs stimulate bFGF expression of retinal Müller cell.Calphostin C inhibits bFGF mRNA increase stimulated by AGEs,and inhibition achieves strongest at concentration 50 nmol/L.Conclusion:AGEs can stimulate bFGF expression of Müller cell to exert the role of angiogenesis .bFGF mRNA expression may be regulated by activation of PKC pathway .

Objective: To investigate the protective effect of zinc on learning and memory of lead-exposed rats and hippocampal SS and SS mRNA neurons. Met hods: Wistar rats were exposed to 6.15 mmol/L lead acetate solution or 6.15 mmol/L lead acetate and 3.10 mmol/L zinc sulfate solution for three months, and the learning and memory ability was studied with Y-maze test. The blood and hippocampal lead concentration were measured with atomic absorption spectrometry and the number of hippocampal SS and SS mRNA positive neur ons were observed with immunohistochemistry and in situ hybridization respectively. Results: Compared with the control and lead-zinc group, the learning and memory ability of lead-exposed rats was significantly decreased(P

Objective: To study the effect of taurine to resist the impact of lead on learning and memory. Methods: ABC immunohistochemistry method was used to study the quantity change of nNOS positive neuron in hippocampus of the rats which were fed with distinct dosage of lead acetate in drinking water (0.02, 0.2 g/L) and distinct dosage of taurine (5,10 g/kg) in feed. Rusults: Taurine could increase nNOS positive neuron quantity obviously in hippocampus of rats induced by lead lesion. Conclusion: Taurine could resist impact of lead on learning and memory obviously.

BACKGROUND:Autologous split-thickness skin grafting is the main therapy for burn repair at functional sites, which has achieved certain effects, but there are stil some deficiencies, such as poor texture, stiffness and poor toughness, as wel as severer hyperplasia that is easy to result in contracture deformity and poor functional recovery. OBJECTIVE: To analyze the clinical efficacy of skin co-transplantation on burn repair at functional sites. METHODS:Sixty patients with burns at functional sites (n=84) were randomized into two groups: co-transplantation of acelular dermis and autologous split-thickness skin in experimental group and autologous split-thickness skin graft in control group. Survival rate of skin flap and rate of secondary operation were compared between two groups. At 1 month after transplantation, Vancouver Scar Scale was used to assess skin color, thickness, blood vessel distribution and flexibility, and meanwhile, the severity of scar was determined. RESULTS AND CONCLUSION:The survival rate of skin flap was significantly higher in the experimental group than the control group (93%vs. 70%,P < 0.05), and the rate of secondary operation was significantly lower in the experimental group compared with the control group (0vs. 13%,P < 0.05). At 1 month after transplantation, scores on the skin color, thickness, blood vessel distribution and flexibility were al lower in the experimental group than the control group (P < 0.05), but the incidence of mild hyperplasia in the experimental group was significantly higher than that in the control group (52% vs. 29%,P < 0.05). These findings indicate that co-transplantation of acelular alogeneic dermis and autologous split-thickness skin for burn repair at functional sites can effectively enhance the survival rate of skin flap, reduce the rate of secondary operation, contribute to wound healing and reduce the severity of hyperplasia.

Persistant infection of high-risk human papillomavirus (HPV) is the primary cause leading to cervical cancer, which is ranked as second cancer threatening the health of women following breast cancer.Development of HPV vaccine is very important because there is no effective therapeutics for cervical cancer.Three currently licensed HPV vaccines based on major capsid protein L1 in the foreign market confered good safety and efficacy in clinical trials, but the current price is expensive due to high cost, which limits the wide application in developing countries.So far, the vaccines have not been launced in China market.Here, we review the progress and the current status of the HPV vaccine, which will attract the readers’ interest on the forthcoming emergence of HPV vaccine in China.

Objective To study the inactivation kinetics and injury mechanism of Escherichia coli under UV disinfection in drinking water.Methods The inactivation kinetics of E.coli ATCC 25922 was determined by plate count methods in the UV disinfection experiment.The morphology,cell membrane permeabilization and injury of biological macromolecules were observed under a transmission electron microscope (TEM).The rate of ONPG hydrolysis and changes in activities of antioxidant enzymes were observed Raman spectroscopy.Results the changes of damaged cells involved morphological damage such as loss of the structural integrity of the wall and membrane,condensation of cellular material and leakage of significant amounts of cytoplasmic material,a more than four-fold increase of cell membrane permeabilization and damage to the structure of protein,nucleic acids and phospholipid.Conclusion UV disinfection can lead to a multi-target damage.

Objective To establish a mouse model of genital human papillomavirus (HPV) pseudovirion (PsV) transmission and evaluate the protective potency of HPV16 VLP vaccine.Methods HPV16 PsV with the encapsidated luciferase expressing plasmid Luc were generated from 293FT cells and purified by size-exclusion chromatography.The endpoint titers of HPV16 PsV-Luc were determined on 293FT cells, denoted as TRLU/mL.For in vivo genital challenge, mice were synchronized in a diestrus-like status by a subcutaneous injection with 0.1 μg β-estradiol and then with 3mg DepoProvera after 24 hours.Six hours prior to HPV16 PsV-Luc challenge, deeply anesthetized mice were intravaginally pretreated with 50 μL of 4%nonoxynol-9 ( N-9 ).HPV16 PsV-Luc was thoroughly mixed with 20 μL solution containing 4%carboxymethylcellulose ( CMC ) and intravaginally instilled using a positive-displacement pipette.Forty-eight hours after PsV-Luc challenge, mice were anesthetized and D luciferin was intravaginally instilled.After 3 minites, bioluminescence was measured with a cryogenically cooled Xenogen IVIS camera system.Then,the murine genital challenge model was used to determine the potency that HPV16 VLP vaccine is efficient at preventing HPV infection.Results HPV16 PsV-Luc was generated and purified from 293FT cells.HPV16 PsV-Luc was verified to containe L1 and L2 protein by Western blot.HPV 16 PsV-Luc successfully infected vaginal epithelial cells of mouse and the murine genital challenge model was established.To achieve consistent bioluminescence, the minimal dose of HPV16 PsV-Luc was 1.7 ×104 TRLU.The protective potency of HPV16 VLP vaccine was shown using this murine model.Our data showed that immune serum with over neutralizing antibody titer of 256-fold was sufficient to confer protection against HPV PsV genital infection .Conclusion The murine genital challenge model of HPV16 was successfully established, and the model is used to evaluate the potency of HPV16 VLP vaccine against in vivo genital HPV16 PsV challenge.Our model will benefit for the investigation of HPV neutralization and the potency evaluation of the HPV vaccine .

Objective:To study the effects of T-2 toxin on expression of fibroblast growth factor 8 (FGF8) and fibroblast growth factor receptor 3 (FGFR3) in articular cartilage and subchondral marrow of rats under low selenium condition, and to explore the mechanism of deep cartilage injury and secondary complications in Kaschin-Beck disease (KBD).Methods:Twenty-four healthy male SD rats weighted 60 - 80 g were selected, they were divided into conventional feed group (selenium content of 101.5 μg/kg) and low-selenium feed group (selenium content of 1.1 μg/kg) by random number table method, with 12 rats in each group. After 30 days of feeding, the conventional feed group was further divided into control group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium feed group was further divided into low-selenium group and low-selenium+ T-2 toxin group, with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the knee cartilage with cancellous bone was taken. Pathological changes of knee cartilage were observed by HE staining. Immunohistochemical method was used to detect the expression of FGF8 and FGFR3 in cartilage and subchondral marrow of knee joint, positive expression rates of FGF8 and FGFR3 in articular cartilage were calculated, and the integrated optical density (IOD) values of FGF8 and FGFR3 positive expression in subchondral marrow were analyzed by Image-Pro Plus 6.0 software. Results:Under light microscope, chondrocytes in low-selenium+ T-2 toxin group were sparse, and empty chondrocytes in the deep and middle layers of articular cartilage increased, and chondrocytes died and became red cell shadows. The extracellular matrix dissolved and was slightly stained in deep region, turning into necrotic and unstructurized areas. Proliferating granulation tissue was visible nearby. The positive expression rate of FGF8 in articular cartilage of rats in low-selenium+ T-2 toxin group [(88.61 ± 10.97)%] was higher than that in control, low-selenium and T-2 toxin groups [(10.35 ± 2.48)%, (19.26 ± 3.08)%, (58.89 ± 9.29)%, P < 0.05]; IOD value of FGF8 positive expression in subchondral marrow [(16.73 ± 1.72) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(1.20 ± 0.41) × 10 6, (4.33 ± 0.97) × 10 6, (12.80 ± 1.12) × 10 6, P < 0.05]. The positive expression rate of FGFR3 in articular cartilage of rats in low-selenium+ T-2 toxin group [(89.76 ± 8.59)%] was higher than that in control, low-selenium and T-2 toxin groups [(13.18 ± 2.25)%, (21.15 ± 2.33)%, (32.55 ± 6.72)%, P < 0.05]; IOD value of FGFR3 positive expression in subchondral marrow [(16.50 ± 5.36) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(7.58 ± 1.02) × 10 6, (10.73 ± 7.13) × 10 6, (9.83 ± 5.63) × 10 6, P < 0.05]. Conclusion:Under low selenium condition, T-2 toxin changes expression of FGF8 and FGFR3 in deep chondrocytes of articular cartilage and subchondral marrow in rats, elevated expression of FGF8 and FGFR3 may be involved in the occurrence and development of secondary changes in KBD.

Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID₅₀, respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 μg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 μg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10⁴ and 1×10⁵. The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.

Objective:To evaluate the characteristics of not just right experiences (NJREs) with high and low obsessive-compulsive beliefs in obsessive-compulsive disorder (OCD) patients, and to analyze their relationship with OCD symptoms.Methods:A total of 142 patients with OCD and 51 patients with anxiety disorders were included in the outpatient department of Beijing Anding Hospital.The Yale-Brown obsessive-compulsive scale (Y-BOCS), obsessive belief questionnaire (OBQ-44), revised version of the not just right experience questionnaire (NJRE-Q-R), Beck depression inventory (BDI), and Beck anxiety inventory (BAI) were used to evaluate clinical symptoms in patients with OCD and anxiety disorders. Cluster analysis was used to divide OCD into high obsessive beliefs group (OCD-H, n=77) and low obsessive beliefs group (OCD-L, n=65).The SPSS 23.0 software was used for data analysis.Chi-square test and one-way analysis of variance were used to compare the demographic and clinical data of OCD-H, OCD-L, and anxiety groups. Multivariate Logistic regression analysis was used to analyze the risk factors for OCD, and multiple linear regression analysis was used to analyze the influencing factors of obsessive compulsive symptoms in the OCD-H and OCD-L groups, respectively. Results:There were significant differences in the scores of Y-BOCS((23.14±6.60), (17.77±6.48), (8.70±6.80)), OBQ-44((162.69±33.15), (86.54±19.09), (103.12±45.67)), NJRE-Q-R checklist scale((4.58±2.61), (2.92±2.15), (2.86±1.72)), NJRE-Q-R severity scale((32.86±8.97), (24.75±9.71), (18.86±8.51)) among OCD-H, OCD-L, and anxiety groups( F=68.87, 102.29, 11.06, 32.01, all P<0.001).Multivariate Logistic regression analysis showed the NJRE-Q-R severity score was a risk factor for OCD( B=0.124, OR=1.132, 95% CI=1.071-1.197, P<0.001).For the OCD-H group, Y-BOCS scores were influenced by disease duration, BDI scores, OBQ-44 factor 1 and factor 3 scores( B=0.020, 0.201, 0.133, 0.126, all P<0.05). For the OCD-L group, the Y-BOCS scores were influenced by the scores of BDI and the checklist and severity score of NJRE-Q-R( B=0.265, 0.852, 0.191, all P<0.05). Conclusions:NJREs are prevalent in OCD patients regardless of the level of OCD beliefs, with higher degrees than anxiety disorders. NJREs are a risk factor for OCD, especially for patients with low obsessive compulsive beliefs.NJREs may be a potential cause and intervention target for OCD patients especially with low OCD beliefs.