Background The effects of Toll-like receptor 2 (TLR2) in grafting-related immune diseases have attracted more and more attention.Blocking TLR2 signal pathway can extend the survival time of heart and kidney grafts.However, the effects of anti-TLR2 monoclonal antibody on corneal graft have not been confirmed.Objective This study was to investigate the influence of anti-TLR2 monoclonal antibody on corneal graft survival in the rats received penetrating keratoplasty (PKP).Methods Allograft corneal transplantation was performed on the right eyes of 24 SPF female Wistar rats to establish PKP models,with 12 SD rats as donors.The model eyes were randomized into the TLR2 monoclonal antibody group and the model group.Anti-TLR2 monoclonal antibody of 15 μg/30 μl was subconjunctivally injected on day 0,2,4,6 and 8 following the modeling in the TLR2 monoclonal antibody group,and equal amount of normal saline was injected in the same way in the model group.The edema,transparency and neovascularization were observed under the slit lamp microscope after surgery, and rejection index (RI) was scored based on the criteria of Holland.Corneal tissue sections of the rats were prepared for the histopathological examination on day 9 and 15 after operation.The research protocol was approved by the Southern Medical University Ethics Committee.Results Mild corneal edema was found in the two groups 1-4 days after operation.A lot of new blood vessels, edema and opacification of corneas were seen in the model group 9-14 days after operation,but in the TLR2 monoclonal antibody group,corneal opacification was found 15 days after operation.The RI scores were significantly higher in the model group than those in the TLR2 monoclonal antibody group 5,9,15 days after operation (t=4.183,4.954,13.506;all at P＜0.05).The survival time in the TLR2 monoclonal antibody group was 15.5 days,with the 95％ confidence interval (CI) 14.9-16.1;while that in the model group was 9.5 days,with the 95％ CI 8.7-10.3, showing a significant difference between the two groups (Z =12.728,P =0.001).The corneal histopathological examination revealed that corneal stromal edema,infiltration of inflammatory cells and vascular lumen were more prominent 9 and 15 days after operation in the model group than those in the TLR2 monoclonal antibody group.Conclusions Anti-TLR2 monoclonal antibody can inhibit inflammatory response after allograft corneal transplantation and therefore extend the survival time of graft in rats.

Objective To compare the efficacy of photodynamic therapy (PDT) alone or in combined with ranibizumab versus ranibizumab monotherapy (intravitreal injection,IVR) in patients with polypoidal choroidal vasculopathy (PCV).Methods 80 eyes of 72 patients with PCV were enrolled into this retrospective and comparative study according to their therapeutic plan.30 eyes of 28 patients,28 eyes of 30 patients and 22 eyes of 21 patients were divided into PDT group,ranibizumab 0.5 mg group (IVR group) or the combination group,respectively.The patients with PCV were diagnosed according to clinical symptoms,optical coherence tomography (OCT) and fluorescent indocyanine green angiography (ICGA).The baseline best-corrected visual acuity (BCVA) before treatment was more than 0.05,and there was no retinal fibrosis and scar for all patients.There was no statistical difference of age (F=0.187),gender (x2 =0.423),average BCVA (F=1.120) and central retinal thickness (CRT) (F=0.431) among three groups (P＞0.05).They had not received any treatment before.Patients received verteporfin PDT in PDT group,3 consecutive monthly IVRs starting day 1 in IVR group,and 3 IVRs after 3 days,1 month,2 months of PDT starting day 1 in combination group.Re-treatment was considered 3 months later if the follow up shown no changes in fundus photography,OCT and ICGA.The average follow-up time was 19 months.BCVA at baseline and follow-up visit at 1,3,6,12 months was measured,and the proportion of patients with ICGA-assessed complete regression of polyps at month 6 was recorded as primary outcome.The CRT was measured at baseline and 6 months as secondary outcome.Results There were significant difference of BCVA at 1,3,6 and 12 months among three groups(F=5.480,5.249,3.222,4.711;P＜0.05).The average BCVA was significantly better at 1,3,6,12 month than that at baseline(t =-6.632,-4.127,-3.904,-4.494;P＜ 0.05) in combination group,and was significantly better at 3,6,12 months than that at baseline (t=-5.636,-3.039,-3.833;P＜0.05) in IVR group.However there was no significant difference of the average BCVA in PDT group between follow-up at 1,3,6,l 2 months and baseline (t=1.973,0.102,-0.100,-0.761;P＞0.05).The proportion of patients with complete regression of polyps at 6 months was higher in PDT (76.7％) or combination group (68.2％) than IVR group (35.7％) (x2=0.003,0.025;P＜0.05).There was no significant difference of CRT among 3 groups at baseline (P=0.651).The mean CRT decreased in all 3 treatment groups over 6 months (t=5.120,3.635,5.253;P＜0.05),but there was no significant difference of CRT among 3 groups (F=1.293,P＞ 0.05).Conclusions Three therapies could effectively decrease CRT.IVR or IVR combined with PDT are both more effective than PDT therapy to improve vision of PCV patients.PDT or PDT combined with IVR was superior to IVR pnly in achieving complete regression of polyps in 6 months in PCV patients.

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.

Objective To reduce the rate of unscheduled nasogastric extubation for the patients in neurology department.Methods A nursing care team was set up for a thematic activity to reduce the rate of nasogastric tube unscheduled extubation.The team analysed the situations of 164 patients with nasogastric intubation in the neurology department,specifying the problems,deciding their aims and working out the strategies.Result The rate of unscheduled nasogastric extubation decreased from 19.63％ to 2.12％ and the rate of nursing omission decreased from 9.93％ to 2.59％ (P＜0.05).Conclusion The improvement activity of nursing projects can effectively reduce the rate of unscheduled nasogastric extubation and improve the quality of nursing care.

Objective To explore the role of type Ⅶ collagen (COL7A1) gene in the pathogenesis of pretibial dominant dystrophic epidermolysis bullosa (DDEB-Pt).Methods Peripheral blood samples were obtained from a sporadic Chinese patient of Han nationality with DDEB-Pt,his parents and 100 healthy human controls.A modified salting-out method was used to extract genomic DNA from the blood samples,and PCR was performed to amplify 118 exons of the COL7A1 gene followed by DNA sequencing.Results A G→A mutation was identified at position 6109 (G6109A) in exon 78 of the COL7A1 gene in this patient,which caused a change from GCT to ACT at codon 2037 in the triple helix region,and resulted in the substitution of glycine (Gly) by arginine (Arg) (p.Gly2037Arg).Conclusion A novel glycine substitution mutation was identified in the COL7A1 gene in the patient with DDEB-Pt,which may be a pathogenic mutation.

Objective To study the situation of missing detection of HBsAg negative samples with high value and to explore the ways for reducing the missing detection .Methods According to the document EP15-A2 issued by the Clinical and Laboratory Standards Institute(CLSI) ,the precision and accuracy of the HBsAg quantitative detection by the ARCHITECT I 2000 were verified for ensuring the instrument conditions conforming to the test requirements .The 1 130 samples with the ratio of the sample absor-bance value to critical value(S/CO) in the range 0 .30-0 .99 in the HBsAg qualitative results by ELISA were collected during the study period and re-tested .After re-testing ,the quantitative HbsAg of 387 samples ,in which the ratio of S/CO was still in the range 0 .30-0 .99 ,were detected by the ARCHITECT I2000 and the detection results were taken as the reference .The missing detection situation of HBsAg negative samples with high value was researched .The ROC curve was adopted to reassess the cutoff value of negative HBsAg with positive HbcAb for searching better diagnostic performance .Results The HBsAg detection results by the i2000SR had good repeatability with total imprecision of less than 9 .88% and the deviation value with calibrator was less than 3 .91% ,which conformed to the clinical testing requirements .This study found that the HBsAg samples with the S/CO ratio of 0 .30-0 .99 had the significant distribution differences in the different batches of test (P<0 .05) .In the study period ,261 cases of HBsAg missing detection occurred ;then the critical value of HBsAg qualitative results detected by ELISA in the HbcAb positive group was re-determined as 0 .51 by using the ROC curve ,among them 239 cases were rejudged as positive .Conclusion Increasing the repeatability of HBsAg qualitative detection by ELISA and establishing the new positive judgement value of ELISA for qualita-tive HBsAg in the HbcAb positive population as 0 .51 possesses better diagnostic performance for detection the HBsAg samples with the S/CO ratio of 0 .30-0 .99 .

Objective To investigate the clinical and genetic characteristics of a progressive symmetric erythrokeratoderma (PSEK).Methods The clinical and genetic characteristics of a PSEK were compared and analyzed with clinical collection and review of 17 PSEK families reported in China since 1980.Results PSEK was consistent with autosomal dominant inheritance.PSEK had the feature with incomplete penetrance and variable expression.The clinical features included hyperkeratotic plaques with distinct border and strikingly symmetric distribution pattern on the extremities.Part of patients was extended to other areas of the body.The onset of the disease commonly started in infancy or childhood.The general health condition was not affected.PSEK might be associated with other clinical symptoms.The diseased potential increased in the family of consanguineous marriage.Conclusions PSEK has genetic heterogeneity.Its causative genes have not been determined.Further studies are needed.

Objective To measure the changes of total factor productivity (TFP)of Zhejiang medical facilities for decision makers to promote their service efficiency.Methods Collecting the panel data of 2005-2010(four input indexes and five output indexes)from medical facilities in Zhejiang province and measuring with Malmquist index of DEA programs of DEAP2.1.Results The average annual TFP growth rate in the period is 2.0％.A further decomposition found that the average annual growth rate of technology progress is 1.8 ％,while that of technical efficiency and pure technical efficiency is only 0.2％respectively.In the meantime,no scale efficiency growth was found.Conclusion The average annual growth of total factor productivity of Zhejiang medical institutions is substantially low in the period,with technology recession found as well.To maximize productivity of the medical sector,the allocation and internal management should be strengthened to stimulate technical efficiency and scale efficiency while encouraging technology innovation.

Objective To analyze the characteristics of genotyping and gene polymorphism of Neisseria gonorrhoeae(N.go) with azithromycin(AZM)-resistance(AZM-R) and decreased susceptibility to ceftriaxone(CROD).Methods The minimum inhibitory concentration(MIC) of AZM and CRO were determined.AZM-R isolates were detected for mutations in 23S rRNA,mtrR and penA genes.Genotypes were analyzed by using N.go multi-antigen sequence typing(NG-MAST).Results All total of 485 isolates of N.go were detected.77(15.9%) strains were AZM-R(MIC≥1 mg/L),including 33(6.8%) isolates of AZM low-level resistant(AZM-LLR,MIC=1 mg/L) strains and 44(9.1%) isolates of AZM middle-level resistant(AZM-MLR,MIC≥2 mg/L) strains.There were more CROD(MIC≥0.125 mg/L) strains in AZM-MLR isolates(43.2%),compared with those in AZM-LLR isolates(18.2%,P<0.05).The detected rates of 23S rRNA,mtrR,penA single or combined mutations were without significant differences between AZM-LLR isolates and AZM-MLR isolates(P>0.05).Similar results were found between combined AZM-LLR/CROD isolates and combined AZM-MLR/CROD isolates(P>0.05).No mutation of A2059G and AZM high-level resistant(AZM-HLR,MIC≥256 mg/L) isolate were found.Among 77 AZM-R isolates,67 sequence types(ST) were identified by NG-MAST,of which 30 types were novel.Most ST were represented by a single isolate.Conclusion AZM-R and CROD isolates,presented in this area,might be deserved continuous surveillance to identify the mechanism of concurrent resistance.

Objective To investigate the role of regulatory T (Treg) / T helper type 17 (Th17) cells in the pathogenesis of chronic spontaneous urticaria (CSU).Methods Eighty-nine patients with CSU were enrolled in this study,including 48 in active stage and 41 in remission stage.Forty-eight health check-up examinees,who were collected from the community hospitals in Guangzhou city,served as the healthy controls.Fluorescence-based realtime quantitative PCR was performed to determine the expression of transcription factors FOXP3 and RORγt in PBMCs from these subjects.Results Compared with the patients with CSU in remission stage and healthy controls,the patients in active stage showed a significantly higher level of FOXP3 mRNA (0.57 ± 0.19 vs.0.11 ± 0.21 and 0.13 ± 0.23,both P ＜ 0.05),but a significantly lower level of RORγt mRNA (0.43 ± 0.39 vs.0.89 ± 0.40 and 0.87 ± 0.43,both P ＜ 0.05).Conclusions The expression of Treg cell regulator FOXP3 increases,while the expression of Th17 cell regulator RORγt decreases in patients with CSU,suggesting that the imbalance between Treg and Th17 cells induced by the interaction between FOXP3 and RORγt may be involved in the pathogenesis of CSU.