Background Cytokines play a crucial role in mediating immune tolerance or immune rejection of corneal transplantation.However,the study on the expression of cytokines in corneal graft is seldom.ObjectiveThe purpose of the study is to investigate the change of cytokine expression in allografts at different time points after corneal transplantation.Methods BALB/c and C57BL/6 mice aged 6 to 8-week old were used to establish autologous and allografts keratoplasty models.BALB/c mouse was use as donor and receipt in autologous group,and the cornea of C57BL/6 mouse was used to graft on the BALB/c mouse in the allografts group.The graft inflammation was clinically scored,and graft inflammatory scores of ≥5 or opaciflcation scores of ≥2 were identified as rejection.BALB/c mice were randomized into normal control group(3 mice)and allografts group(15 mice).Reverse transcriptPCR (RT-PCR) was used to detect the expression of interleukin-4 (IL-4) mRNA,interferon-γ (IFN-γ) mRNA,IL-10mRNA and tumor necrosis factor(TNF-α) mRNA in graft 6 hours and 1 day,3,7,14 days after operation.Results The corneal graft opacification score was ＜2 and inflammatory score was ＜5 in the 10 mice with autologous keratoplasty until 60 days with the survival rate 100％.The edema,opacification and new blood vessel were seen in the BALB/c mice received allografts keratoplasty.The inflammatory score was ≥ 5 and the opacification score was ≥2 24 days after surgery with the rejection rate 100％,in the allografts group,and the graft survival time was (17.80±4.66)days.RT-PCR showed that IL-4 and IFN-γ were positively expressed,and IL-10 and TNF-α were absently expressed in normal mouse cornea.In the allografts keratoplasty mice,positive responses for IL-4,IFN-γ and IL-10 were found in 6hours after operation,but TNF-α was absent.From 1 day through 3 days after operation,the expressions of IL-4,IFN-γand TNF-α were enhanced but IL-10 was disappeared in the graft.IL-10,IFN-γ and TNF-α were expressed till the 7th day,but on the 14th day,only IL-10 was detected in graft in the allografts keratoplasty mice.Conclusions TNF-αis a main factor among the variety of cytokines that may influent corneal allograft rejection locally.

Objective To observe the effects of fetal bovine serum(FBS) on differentiation of human neural stem cells (NSCs). Methods The effects of FBS with different concentrations on differentiation of human fetal NSCs were observed by cell culture, immunocytochemistry and flow cytometry. Results Human fetal NSCs could be induced to differentiate mainly three types of nerve system cells(neuron, astrocyte and oligodentrocyte). There were 80%～90% astrocytes of differentiated cells from human fetal NSCs with the concentration of 15% FBS induced. Conclusion Concentration dependent FBS in culture medium may have effect on the ratio of neurons to glial cells differentiated from human NSCs in vitro .

Objective To identify and investigate B(A)02 allele in a patient. Methods Serological tests were performed with standard serological methods in a patient with B(A)02 allele. DNA sequences of all seven exons and exon-intron boundaries of ABO gene were analyzed by polymerase chain reaction (PCR), direct DNA sequencing and sequencing after gene cloning. In order to analyze the allele, PyMOL software was used to establish 3D model of Glycosyltransferases B (GTB). Results The serological results showed the characteristics of B(A) phenotype. DNA analysis revealed that ABO gene of the individual was heterozygous of B(A)02/O01 allele. 700C>G mutation was identified in B101 allele, which resulted in the amino acid substitution P234A in GTB. Through the analysis of the 3D structure of GTB, it was speculated that the P234A replacement affected the intermolecular forces of the 234 amino acid and Met-266, thus changed the conformation of the donor-binding pocket of GTB,that made GTB capable of recognizing and tranferring the GalNac to the H antigen, which can lead to the formation of the weak A antigen on membrane of red blood cells. Conclusion The P234A replacement can affect the spatial conformation of the specific recognition region conformed by Met-266 and Ala-268 residues, which leads to the antigenicity change of the ABO blood group.

Objective To compare the effects of adjustable pressure drainage device, negative pressure ball drainage and drainage bag drainage on patients with the total hip arthroplasty. Methods The silicone drainage ball with negative pressure, the urine collection bag, the infusion set′s regulator were used to make adjustable pressure drainage device. 120 cases of total hip arthroplasties from January to December in 2015 were chosed and divided randomly into 3 groups based on the sequence of operation notifications. In 40 cases, drainage bag were used (control group 1), 40 cases with negative pressure ball drainage (control group 2) and 40 cases with adjustable pressure drainage device (experimental group). Total postoperative bleeding, the blood transfusion amount, swelling (the thickness of hematoma) and the nursing workload of the three groups were compared. Results The total postoperative bleeding, the blood transfusion amount, the thickness of hematoma and the nursing workload in the experimental group were (775.1±130.5) ml, (3.5±1.3) mm, (180.2±29.9) ml, (34.5±5.2) min. The total postoperative bleeding, the blood transfusion amount, the thickness of hematoma and the nursing workload in the control group 1 were (889.4 ± 160.8) ml, (5.2 ± 1.1) mm, (285.9 ± 30.4) ml, (40.6 ± 7.4) min and in the control group 2 were (876.2±156.7) ml, (4.3±1.2) mm,(208.3±33.3) ml, (50.3±8.5) min. Compared the data of the experimental group and the control group 1, the difference was of statistically significance (t=3.49-15.68, P < 0.01). Compared the data of the experimental group and the control group 2, the difference was of statistically significance (t=2.86-10.03, P<0.01). Conclusions The effects of adjustable pressure drainage device are better than the drainage bag and negative pressure drainage ball. It can effectively reduce the postoperative bleeding and the swelling degree, and bring a lot of relief to the patients and lots of convenience to the nursing work. The method is affordable, reliable and safe which should be promoted in the primary hospital.

Objective To explore the expression and significance of signal transducer and activator of transcription 3 (Stat 3), glucose transporter protein 1 (GluT-1) and proliferation cell nuclear antigen (PC NA) in lesions of condylomata acuminata (CA). Methods SP immunohistochemistry method was used to measure the expression of Stat 3, GluT-1 and PCNA in tissue samples from 40 cases of CA and 20 normal skin controls. Results The positivity rates of Stat 3, GluT-1 and PCNA were 85.0% (34/40), 87.5% (35/40) and 85.0%(34/40), respectively in CA tissue, 35.0% (7/20), 30.0% (6/20)and 55.0% (11/20),respectively in the control tissue; statistical difference was observed in these rates between the two groups (all P ＜ 0.05). The expression intensity of Stat 3, GluT-1 and PCNA was also higher in CA tissue than that in the controls. In addition, the expression intensity of PCNA was correlated with that of Stat 3 and GluT-1in CA tissue (both P＜ 0.05). Conclusions There is an overexpression of Star 3, GluT-1 and PCNA in CA tissue, and the overexpression of Stat 3 and GluT-1 may be associated with the over-proliferation of CA tissue.

Objective To investigate the expressions and effects of EGR-1, p53, p16, Cyclin D1 in Nasopharyngeal Carcinoma (NPC). Methods Expressions of EGR-1, p53, p16, and Cyclin D1 protein in paraffin-embedded specimens of 29 chronic nasopharyngitis, 45 NPC and 24 lymph-node metastasis were studied by immunohistochemical method (SABC). Results The positive rates of EGR-1, p16, and Cyclin D1 protein expressions in chronic nasopharyngitis were 44.83 %(13/29), 89.66 %(26/29) and 24.14 %(7/29) respectively, but no p53 protein expression. In NPC, the positive expression rates of the four proteins were 22.22 %(10/45), 91.11 %(41/45), 53.33 %(24/45) and 66.67 %(30/45), with significant difference compared with chronic nasopharyngitis respectively; and in lymph nodes metastasis were 16.67 %(4/24), 79.17 %(19/24), 70.83 %(17/24) and 75.00 %(18/24) with significant difference compared with those of chronic nasopharyngitis and NPC respectively except for p16 protein. Spearman correlation analysis showed that there were no significant correlation between the expression of EGR-1 and p53, p16, Cyclin D1 respectively both in NPC and in lymph nodes metastasis. In NPC, the expression of p53 showed a positive correlation with the expression of p16; and in lymph nodes metastasis the expression of Cyclin D1 showed positive correlation with the expressions of p16 and p53. Conclusions The expression of EGR-1 protein was significantly decreased both in NPC and in lymph-node metastasis, which suggested that Egr-1 gene may be a tumor suppressor gene and have an important effect in the pathogenesis and the progression of NPC. The overexpression of p53 and Cyclin D1 also have an important role in NPC, but p16 probably have no biological significance in NPC.

Objective To investigate the effects of Stichopus variegatus mucopolysaccharide on plasma nitric oxide (NO) and serum endothelin(ET) after angioplasty in rabbits. Methods 50 New Zealand rabbits were randomly divided into four groups: Stichopus group, simvastatin group, model group and normal group. Endothelium of iliac arteries in Stichopus group, simvastatin group and model group were denuded by balloon catheter and the rabbits were fed with 2%cholesterol, 3%lard and 3%yolk mixed forage for six weeks, and then, atherosclerotic stenosis was showed by iliac angiography.When transluminal balloon angioplasty was done, from then on the feeds was changed from high cholesterol to ordinary forage. Stichopus (0.5g?kg -1?d -1) and simvastatin (0.5mg?kg -1?d -1) were given by gastric canal. While NaCl solution (0.9%) was given in model group. All rabbits were fed in separate cages and free drinking. After four weeks since transluminal balloon angioplasty, blood was collected from carotid artery to measure the changes of concentrations of endothelin(ET) and nitric oxide(NO). Examination of pathology by optical and electron microscope were done at same time. Results After four weeks since transluminal angioplasty, the concentration of plasm ET in model group increased significantly compared with the normal group(114.86?24.89 ng?L -1 vs 75.62?9.68 ng?L -1) and serum NO decreased significantly(37.50? 12.79?mol?L -1 vs 106.37?18.01 ?mol?L -1).The concentration of plasm ET in model group was higher than that in the simvastatin groups and Stichopus groups(114.86?24.89 ng?L -1 , 81.08?14.71 ng?L -1 and 83.34?11.41 ng?L -1 ,respectively),and the concentration of serum NO decreased obviously(95.63?8.62 ?mol?L -1,98.80?9.06 ?mol?L -1 and 37.50?12.79 ?mol?L -1,respectively). Conclusion After iliac arterial TA the concentrations of plasm ET were increased and the serum NO were decreased, the endothelial function fell into disorder. The Stichopus variegatus mucopolysaccharide had the effects of improving the endothelial function after angioplasty in rabbits by adjustment of plasm ET and serum NO.

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