Background Streptozotocin (STZ)-induced type 1 diabetic model is an acceptable model and is often used to the study of diabetic retinopathy (DR).However,the model is often established using retinal digest preparation in vitro in albino rats,and therefore it is difficult to evaluate the change of retinal vessels by fundus fluorescein angiography (FFA) in vivo.Recently,pigmented rats are being used as the DR model.But the study on the comparison between albino rat model and pigmented rat model is seldom.Objective This study was to observe and compare the manifestations of FFA and retinal digest preparation in early diabetic pigmented diabetic rat and albino diabetic rat,and to select the right model of DR using FFA.Methods Type 1 diabetic models were induced in 15 six-week-old health male BN rats and 15 six-week-old health male SD rats by the injection of STZ (55 mg/kg) via tail vein.The models were thought to be successful in the rats with the blood glucose level ≥ 16.7 mmol/L.The right eyes of the rats were extracted 6 weeks after injection of STZ.Lens were examined by slit lamp microscope.Retinal vascular changes were examined by fundus photography,FFA and periodic acid Schiff staining of retinal digest preparation.The manifestations of FFA and retinal digest preparation were contrasted between BN rats and SD rats.The number of eyes with lens opacification was compared by Chi-square test and the ratio of vascular endothelial cells and perithelial cells (E/P) was compared between BN rats and SD rats.The use and care of experimental animals complied the Statement of Ethic Committee of Tianjin Medicine University.Results Six weeks after injection of STZ,11 BN rats and 10SD rats were included in this study.The blood levels were (24.73±2.98) mmol/L and (22.36±3.65) mmol/L in BN rats and SD rats,respectively,without significant difference between the 2 types of rats (t =7.873,P＞0.05).Lens opacification occurred in 6 BN rats and in 5 SD rats (P=0.717).FFA showed the clear retinal vascular under the brown background.Evident vessel disorder and fluorescence leakage like background stage of DR were seen in 9 eyes.However,in the SD rats,retinal vessel abnormality could not exhibited owing to the interference of choroid fluorescent light from choroidal vessels and vortex vein.Retinal digest preparation exhibited that unevenness of vessel diameter,decrease of perithelial cells and increase of endothelial cells in both types of rats.The E/P values were 11.50±3.68in the BN rats and 12.86±3.94 in the SD rats,without significant difference between them (t=9.785,P＞0.05).Conclusions The abnormality of fundus vascular morphology can be better displayed in pigmented diabetic rats than albino rats by FFA in vivo.

Objective Through smdying the distribution of the Sappan acetic ether extract in transplanted heart of rats,to discuss selectivity of sappan in donor's hearts.Methods Improved Ono model was used to replicate the heart transplantation model of rats.Successfully operated rats were separated into 2 groups:a model group and a sappan group.Distributions of sappan in transplanted heart were observed by HPLC-FPC.Results One kind of original substance.of sappan was detected in transplanted heart sample,but not in un-transplanted hearts.Conclusion The substance of sappan may have selectivity in transplanted heart.

Objective To investigate the protective effect of hypoxia-inducible factor-1α(HIF-1 α) on the neurovascular unit in rats with traumatic brain injury (TBI).Methods The fluid percussion model was applied to induce TBI in rats.A total of 600 rats were divided into sham operation group,TBI group,TBI + HIF-1 α silence group and TBI + control virus group according to the random number table,with 150 rats in each.Virus-mediated HIF-1 α silence gene and control virus were delivered 24 h before the fluid percussion injury.After 3,7 and 14 d,brain injury area and morphological changes in injured region were detected by HE staining,expressions of vascular endothelial cell markers (vWF) and HIF-1 α were detected by Western blot method,and expressions of vascular endothelial growth factor (VEGF),matrix metalloproteinase-9 (MMP-9),tumor necrosis factor-α (TNF-α),interleukin 6 (IL-6) and nuclear factor-κB (NF-κB) in peripheral blood and brain tissue were detected by ELISA method.Rat neural function was dynamically assessed using the modified neurological severity score (mNSS).Results (1) Brain injury area and edema area in TBI + HIF-1 α silence group were higher than those in TBI group at all time points (P ＜ 0.05).(2) Compared with sham operation group and TBI + control virus group,expression of HIF-1α in TBI group gradually increased and remained high at 7 and 14 d postinjury (P ＜ 0.05).Compared with TBI group,expression of vWF in TBI + HIF-1αsilence group decreased at all time points (P ＜ 0.05) and inhibited angiogenesis.(3) TBI + HIF-lα silence group versus TBI group showed remarkably decreased VEGF at all time points,increased expressions of TNF-α,IL-6 and NF-κB at all time point,and increased expression of MMP-9 at 7 and 14 d postinjury (all P ＜0.05).(4) TBI + HIF-1α silence group versus TBI group showed significant difference in mNSS at 7 and 14 d postinjury (all P ＜ 0.05).Conclusions After TBI,high expression of HIF-1αcan facilitate vascular formation and inhibit inflammatory reaction related factor expression,inducing the mitigation of brain edema and brain injury.Therefore,promoting HIF-1α expression may become a new means to improvement of neurovascular function after TBI.

Objective To observe the clinical features and prognosis of endogenous bacterial endophthalmitis (EBE).Methods Ten eyes of 10 patients diagnosed with unilateral EBE were retrospectively reviewed,including 7 males and 3 females.The mean age was 57.6± 10.8 years old.Eight patients were with diabetes and 7 of them were diagnosed over 5 years.There were 3 patients with hepatocirrhosis,1 patient with hypertension,and 1 patient with coronary disease.Nine cases had infectious diseases,including liver abscess (7 cases),pulmonary infection (3 cases),erysipelas (1 case) and perianal abscess (1 case).Seven cases had fever history.Culture and drug sensitive tests for aerobic bacteria,anaerobic bacteria and fungal were performed for 9 eyes using vitreous samples from the procedures of vitrectomy and/or intravitreal injection.All patients were treated with broad-spectrum antibiotics and adjusted for drug use according to microbiological culture and drug sensitivity test results.After the diagnosis was established,vitrectomy combined with lens removal was performed in 5 hours (3 eyes) and 24 hours (5 eyes);Vitreous tamponade of C3F8 (1 eye) and silicone oil (7 eyes) was used;At the end of the operation,0.1 ml vancomycin (1 mg) and 0.1 ml ceftazidime (1 mg) were injected into the vitreous cavity.One eye received intravitreal injection of 0.1 ml vancomycin (1 mg) and 0.1 ml ceftazidime (mg),one eye received evisceration.During the follow up period from 6 to 24 months,visual function,slit lamp and fundus examinations were performed at each office visit.Results All patients complained of blurred vision and 5 patients had ocular pain.The visual acuity was no light perception (3 eyes),light perception (5 eyes);hand motion (1 eye) and 0.1 (1 eye).Corneal edema was found in all 10 eyes;hypopyon in 8 eyes;diffuse vitreous opacity in 10 eyes,including 3 eyes with retinal detachment.For 8 eyes treated by vitrectomy and intravitreal injection,1 eye was eviscerated due to uncontrolled inflammation.The eye treated with intravitreal injection was enucleated for its uncontrolled inflammation.For 9 eyes received vitreous culture and drug testing,8 eyes (88.9％) had positive results,including 5 eyes with Klebsiellar pneumonia,and 1 eye with Staphylococcus aureus,or Streptococcus agalactiae or Enterococcus faecalis respectively.At last office visit,2 eyes were with no light perception;4 eyes were with hand motion;and 1 eye with visual acuity of 0.1.Conclusions Most of the patients with endogenous bacterial endophthalmitis have systemic predisposing factors.Klebsiella pneumoniae is the leading cause of ocular EBE.Vitrectomy combined with intravitreal injection of antibiotics showed efficacy in treating EBE.

Objective To study the effect of simvastatin (SIM) on the expression of neuron specific enoalse (NSE) in rat brain and serum after traumatic brain injury (TBI), and therapeutic effects of SIM on TBI thereof. Methods A total of 90 Sprague-Dwalye (SD) rats aged 8 weeks were randomly divided into sham TBI group, control group and treatment group (n=30). The TBI model was established in control group and treatment group by using Feeney method. Rats in treatment group were fed SIM 10 mg/kg in the evening pre-injury and in every evening post-injury while those in control group were fed the same dose of starch at the same time. Blood samples (3 mL) were collected from carotid atrery in three groups, then rats were sacrificed and brains were collected at different time points (3 h, 12 h, 24 h, 3 d, 7 d and 14 d post-injury). The serum ex-pressions of NSE were detected by ELISA method. The NSE expressions in hippocampal area CA3 were detected with immu-nohistochemistry. Results (1) In control group, the serum NSE level was significantly increased at 3 h after injury, reached the peak at 3 d, and was still higher than that of sham injury group at 14 d. In treatment group, the serum NSE level was in-creased 3 h after injury, reached the peak at 24 h, decreased after 3 d, and was near the sham injury group at 14 d after inju-ry, but was significantly lower than that of control group. (2) Immunohistochemical detection showed that the NSE optical density values in hippocampal area CA3 area were decreased at 3 h after injury in control group. The optical density values reached the lowest level between 3 d to 7 d and were still significantly lower than those of sham injury group at 14 d. In treat-ment group the optical density value was decreased at 3 h after injury, reached the lowest level between 12 h to 24 h and re-bounded significantly at 7 d, then at 14 d up to the level of sham injury group. Conclusion SIM can promote the decrease of serum NSE level in TBI rats and increase the NSE expression of hippocampal neurons of injured side, showing protective effects on neuronal damage after traumatic brain injury.

Abstract: Objective　The method of "molecular docking - molecular dynamics - in vitro pharmacology" was used to screen and optimize small molecule polypeptide compounds that can block PD-1/PD-L1 binding and have high anti-tumor activity. Methods Using the 5-peptide compound database constructed by our team and the three-dimensional structure data of the PD-L1 protein downloaded from the protein database (PDB), the peptide compounds obtained by flexible molecular docking using Molecular Operating Environment software. The molecular dynamics calculation and analysis of the compounds with the highest GWVI/WSA (generalized Born volume integral/weighted surface area) free energy change value (ΔG) were carried out, including the root mean square deviation (RMSD) of the position change of the weight atom and the interaction energy (the sum of Lennard-Jones potential and Coulombic energy). The blocking effect of ligand compounds on PD-1/PD-L1 binding was analyzed by the homogeneous time-resolved fluorescence (HTRF) technique. The co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells was established to explore the effect of compound ligands on the killing effect of T cells on tumors and the effect on the secretion level of IL-2 in the co-culture supernatant. Results The selected polypeptide compounds were analyzed by molecular dynamics, and the binding of RGGHA to RGGHH and PD-L1 was stable. There are many kinds of interactions between RGGHA and PD-L1，which can compete with PD-1 ligands to bind Asp122, Try123 and Lys124 sites of the PD-L1 protein and block PD-1/PD-L1 signal transduction. HTRF experiments showed that the binding inhibition rate of RGGHA to PD-1 and PD-L1 was 58.38%, and that of RGGHH was 42.73%. In addition, we established a co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells to explore the immunostimulatory effect and mechanism of peptides. The results show that RGGHA and RGGHH can significantly increase the secretion level of IL-2, improve the killing ability of T cells against tumor cells and activate the tumor immune microenvironment. Conclusions The study found that the polypeptide compounds RGGHA and RGGHH can effectively block the PD-1/PD-L1 interaction and reactivate the anti-cancer immune response, which can be used as lead compounds for new drug development.

Objective: To investigate the hospital employee satisfaction and trustworthiness in the background of the trusteeship mode and to find out the problems after trusteeship.Methods: Minnesota satisfaction questionnaire (MSQ) and the self-made questionnaire were used to conduct the survey.Results: The overall employee satisfaction (3.80±0.86) and trustworthiness (3.95±0.77)were higher.The highest level of trustworthiness concerned the cultural connotation (84.8%) followed by the management concept (82.8%) for overall satisfaction.The lowest level of employee satisfaction concerned income and workload (53.7%), followed by the working conditions and environment (55.3%).The administrative staff satisfaction was higher compared to that of medical staff (p=0.001), which showed significant statistical differences.Conclusions: The hospital has made some achievements after the trusteeship system reform, however, it still needs improvement and further strengthening in many aspects.The hospital must always keep abreast of the demands of workforce and improve the staff satisfaction so as to promote its continuous and sustainable development.

OBJECTIVETo assess the reporting quality of randomized controlled trials (RCT) related to dentine hypersensitivity which were published between 2000 and 2009 in nineteen Chinese stomatological journals, learn the current status and the influence factors of reporting quality of the dentine hypersensitivity RCT, and determine whether they can provide high quality evidence clinically.

METHODSA handsearching of nineteen Chinese stomatological journals to identify dentine hypersensitivity RCT which were published between 2000 and 2009 and labeled "random". Assess the reporting quality of these RCT by Consolidated Standards of Reporting Trials (CONSORT) statement and then analyze the influence factors of reporting quality by univariate and multivariate analyses.

RESULTSWe identified 61 dentine hypersensitivity RCT, among which 45 belonged to journals of Chinese scientific study statistical source and 16 belonged to journals of non-statistical source. The reporting quality of dentine hypersensitivity RCT was not high at large and the CONSORT score was 8.1 +/- 2.4 on average. Multiple linear regression showed that the score of improved Jadad scale (t=4.656, P=0.000) was the main influence factor of reporting quality of dentine hypersensitivity RCT. The better the intrinsic authenticity was, the higher the reporting quality would be.

CONCLUSIONCurrently, reporting quality of Chinese dentine hypersensitivity RCT needs to be improved. In order to improve the reporting quality of dentine hypersensitivity RCT, experts in stomatology should not only improve experimental design before experiments, but also write papers in accordance with CONSORT statement.

Radiotherapy is a treatment for cancer with undesired by-effects. In order to develop a new radiation protective agent that could reduce the by-effects, we tried to express and purify human cryptochrome 1 (hCRY1). The coding sequence of hCRY1 was inserted into prokaryotic expression plasmid pET28a(+), and this protein was purified from Escherichia coli BL21(DE3) after IPTG induction, ultrasonication, inclusion body dissolution, gradient dialysis, nickel column purification and ultrafiltration. The yield of hCRY1 in 1 L E. coli culture (LB medium) was about 10-15 mg. The radiation protective efficiency of hCRY1 was monitored by detecting X-ray-induced H2A.X foci in HaCaT cells. The results of immunofluorescence show that hCRY1 significantly reduces X-ray stimulated DNA damage response. The apoptosis of HaCaT cell was also detected, and the repression of H2A.X foci formation was not due to hCRY1's cytotoxity. All these data suggest a potential application of recombinant hCRY1 as a protective agent for radiotherapy.
Cryptochromes ; biosynthesis ; Escherichia coli ; Humans ; Plasmids ; Radiation-Protective Agents ; Recombinant Proteins ; biosynthesis

Objective: : The function of B7H3, a member of the B7 family of proteins, in neuroblastoma (NB) remains poorly characterized. Here we examine the expression pattern of B7H3 in clinical NB specimens and characterize the phenotype of B7H3 knock-down in NB cell line. Methods: : Immunohistochemical (IHC) staining was carried out to assess the expression of B7H3 in clinical NB specimens. Survival association was analyzed using five Gene Expression Omnibus (GEO) datasets (GSE85047, GSE45480, GSE62564, GSE16476, GSE49710). Clonogenic survival and flow cytometry were performed after B7H3 knockdown to assess the cellular proliferation and cell survival in vitro. Impact of B7H3 silencing on NB growth was examined in vivo using the SH-SY5Y xenograft model. Results: : On IHC staining, B7H3 was widely expressed in clinical NB specimens. Analysis of the transcriptional profiles of five GEO datasets clinically annotated NB specimens revealed that decreased B7H3 expression was associated with improved overall survival. B7H3 knockdown suppressed the proliferation of the SH-SY5Y NB model in vitro and in vivo. Cell cycle analysis revealed that B7H3 silencing induced G1/S arrest. This arrest was associated with the suppression of E2F1 expression and induction of Rb expression. Conclusion : Our results demonstrate that B7H3 expression correlate with clinical survival in NB patients. Preliminary studies suggest that B7H3 may mediate the G1/S transition.