[Analysis of clinical aesthetic effect of buccal alveolar ridge preservation and connective tissue transplantation with single implant].

PURPOSE: To evaluate the clinical aesthetic effect of buccal alveolar ridge preservation (ARP) and connective tissue transplantation (CTG) in patients who received a single implant. METHODS: Forty-three patients with tooth loss admitted to the Department of Stomatology of Shunde Hospital of Southern Medical University from May 2014 to May 2016 were included in the study. Tooth extraction, ARP, implant implantation, CTG and permanent repair were performed respectively. The incidence of bleeding, depth of probing, marginal bone resorption, and red-white aesthetic effect of implants were evaluated 1 year and 3 years after surgery. The buccal mucosa thickness of implants before, immediately after CTG, 1 year and 3 years after surgery were measured. The patient satisfaction was evaluated by visual analogue scale (VAS) from masticatory function, overall aesthetics, attachment height, and color, respectively. The implant conditions at the third year after surgery were observed, and complications during follow-up were recorded. SPSS 20.0 software package was used for statistical analysis of the data. RESULTS: The follow-up rate in the first year after surgery was 100%, and that in the third year after surgery was 90.70%. One year and 3 years after operation, the aesthetic effect of the implant was satisfactory. At the 3rd year after operation, the scores of the near middle gingival papillary were significantly higher than that at the 1st year after operation (P<0.05). The buccal mucosal thickness of the implant immediately after CTG and 1 year and 3 years after surgery increased significantly compared with that before CTG (P<0.05). The buccal mucosal thickness of the implant increased 1.02 mm (relative stability: 90.12%) 1 year after operation and 1.01 mm (relative stability: 84.31%) 3 years after operation, respectively. The satisfaction scores of the patients on chewing function, overall aesthetics, attachment height and color of the implant immediately after CTG, one year after surgery and 3 years after surgery were all > 8. The 3-year survival rate of the implants was 100%, and the 3-year success rate of the implants was 97.44%. During the follow-up, two patients developed peri-implant mucositis, which was relieved after tooth cleaning, but no complications such as tissue flap necrosis, limited opening and tongue movement disorder occurred. CONCLUSIONS: ARP and CTG have good clinical and aesthetic effects on patients with tooth loss. In three years, the buccal mucosal thickness of the implant can be increased and relatively stable, which is worthy of clinical promotion and application.

A Risk Score System for Myopia Symptom Warning.

Myopia is the leading cause of visual impairments worldwide. Some studies revealed that visual experience in early life affected the final myopia, indicating that environmental factors play an impellent role in the development of myopia. However, risk factors of myopia are still not identified among adolescents in China. A total of 4104 cases of myopia symptom and 3306 emmetropia controls were selected from students in primary and middle schools in Wuhan in 2008. We identified the risk factors associated with myopia symptom by multivariate logistic regression in this cross-sectional study and constructed a risk score system for myopia symptom. The value of the area under the receiver operating characteristic curve (ROC) was 0.735. Furthermore, we followed up 93 students aged 7-9 years for one year and calculated the total points using the score system. We found no significant difference between the final myopia symptom and the results predicted by the total points by pair chi-square test (P>0.05). The score system had a modest ability to estimate the risk factors of myopia symptom. Using this score system, we could identify the students who are at risk of myopia symptom in the future according to their behaviors and environmental factors, and take measures to slow the progress of myopia symptom.

Diaqua-bis-(3-nitro-benzoato-κO)bis-[1H-5-(3-pyrid-yl)-3-(4-pyrid-yl)-1H-1,2,4-triazole-κN]cobalt(II) dihydrate.

In the centrosymmetric title compound, [Co(C(7)H(4)NO(4))(2)(C(12)H(9)N(5))(2)(H(2)O)(2)]·2H(2)O, the Co(II) atom, located on an inversion center, is coordinated by two N atoms [Co-N = 2.155 (3) Å] and four O atoms [Co-O = 2.099 (2)-2.117 (3) Å] in a distorted octa-hedral geometry. Inter-molecular N-H⋯O, O-H⋯N and O-H⋯O hydrogen bonds link the components into a three-dimensional supramolecular framework.

[Construction of the eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 and green fluorescent protein].

OBJECTIVE: To construct an eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and enhanced green fluorescent protein (EGFP) controlled by cytomegalovirus promoter using pIRES-EGFP vector. METHODS: The mtHSP70 gene fragment was amplified by PCR from pVAX-mtHSP70-HSV2gD using specific primers. The PCR product was cloned into the vector pMD 18-T vector, and the correct clone was selected according to DNA sequence analysis. The interested mtHSP70 gene fragment was subcloned into pCMV-IRES-EGFP vector with XhoI and EcoR I digestion. The recombinant plasmid was transfected into mouse melanoma B16 cell line, and the green fluorescent cells were detected by fluorescence microscopy and mtHSP70 expression was detected by Western blotting. RESULTS: The recombinant plasmid obtained was confirmed by enzyme digestion. The transfected mouse melanoma B16 cells exhibited green fluorescence under fluorescence microscopy and expressed mtHSP70 protein as demonstrated by Western blotting. CONCLUSION: The eukaryotic coexpression vector PCMV-mtHSP70-IRES-EGFP has been established to allow further investigation of the role of mtHSP70 gene in tumor immunotherapy.

[Culture and identify the human umbilical vein endothelial cells and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains in the cells].

OBJECTIVE: To study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs. METHODS: HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry. RESULTS: HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%. CONCLUSION: Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.

[Effect of vascular endothelial growth factor 165 gene transfection on repair of bone defect: experiment with rabbits].

OBJECTIVE: To investigate the influence of vascular endothelial growth factor (VEGF) 165 gene transfection on the repair of bone defect. METHODS: 38 New Zealand rabbits underwent resection of a segment 1 cm in length in bilateral radii filled with absorbable gelatin sponge. Dilated solution of the plasmid pcDNA3.1/VEGF165 was injected into the bone defect of one side and normal saline was injected into the contralateral bone defect. 1, 2, 4, 6, 8, and 12 weeks later X ray examination was conducted to observe the repair of bone defect, and then 5 rabbits were killed at each time points to take out the bone defects. HE staining was used to observe the bone repair. The levels of microvessel density (MVD) 1 and 2 weeks after the operation were observed. RT-PCR was used to detect the mRNA expression of VEGF in the bone defect. Based on the results of RT-PCR the tissue mRNA expression of VEGF65 was detected by real-time quantitative polymerase chain reaction (RQ-PCR). RESULTS: X-ray examination showed that there was no significant difference in the wound healing between the two group 1 week after the operation in all rabbits. Some callus could be seen in the experimental group 2 weeks after. Twelve weeks after the operation the reconstruction of bone cortex was completed. Similar process occurred in the control sides but more lately. The MVD level 7 days after of the experimental group was 47.0 +/- 7.5, significantly higher than that of the control group (42.2 +/- 6.4, t = 2.4519, P = 0.0179), and the MVD level 14 days after of the experiment group was 69.1 +/- 5.4, significantly higher than that of the control group (56.1 +/- 6.1, t = 8.0347, P = 0.0000). In the experimental group the mRNA expression amounts of VEGF165 could be found 1 week after, gradually increased and peaked 3 weeks after, then decreased, and became stable 6 weeks after. The mRNA expression amounts of VEGF165 in the control group were lower than those of the experimental group. CONCLUSION: Local application of PcDNA3.1/VEGF(165) vector promotes the expression of VEGF165, and enhances the quantity of the angiogenesis, extra cellular matrix and healing of bone defect.

Effect of vascular endothelial growth factor 165 gene transfection on bone defects and its mRNA expression in rabbits.

BACKGROUND: Gene therapy has been a hot spot in repair of bone defects in recent years. This study aimed to construct a recombinant plasmid pcDNA3.1-VEGF(165), and to observe the effect of vascular endothelial growth factor 165 (VEGF(165)) gene therapy on bone defects in rabbits. METHODS: Total RNA was extracted from rabbit bone tissues. VEGF(165) cDNA fragment was prepared by reverse transcription and the gene was cloned by polymerase chain reaction (PCR). Plasmid pMD18-T/VEGF(165) combined with pcDNA3.1 was cloned to reconstruct pcDNA3.1-VEGF(165) plasmid. Thirty New Zealand white rabbits weighing (2.50 +/- 0.13) kg were used to establish models of bone defects (1 cm in length) of the bilateral radii. The bone defects were repaired with absorbable gelatin sponge. After the operation, physiological sodium chloride solution was injected into the injured site in one of the forelegs of the rabbits as the control group, and pcDNA3.1-VEGF(165) plasmid (0.2 ml, 200 ng) was injected into the opposite foreleg as the experiment groups. At weeks 1, 2, 4, 6, 8, and 12 after the treatments, the bones were examined by X-ray, and the specimens of the bone defects were collected, stained with HE, and observed under a light microscope. The expression of VEGF(165) mRNA was examined by real-time quantitative polymerase chain reaction (RQ-PCR). RESULTS: The pcDNA3.1-VEGF(165) plasmid with a correct sequence was constructed successfully. Postoperative X-ray found no difference between the two groups at week 1. In the experiment group, callus and synostosis were observed after 2 weeks, and osteosis structure was normal at week 12; these phenomena occurred much later in the control group. In the experiment group, HE staining showed a large amount of newly formed blood vessels after 2 weeks, a number of bone trabeculae with osteoblasts proliferation at 4 weeks, and fresh bone cortex and reformed medullary cavity at 12 weeks; whereas in the control group these structures formed in later phases. The VEGF(165) mRNA in the experiment group was expressed at a low level at week 1, reached the peak at weeks 3, and then decreased to a normal level after 6 weeks. CONCLUSIONS: Local use of pcDNA3.1-VEGF(165) plasmid at bone defects can upregulate the expression of VEGF(165) and accelerate the formation of capillaries and the repair of bone defects. Angiogenesis and osteogenesis can be promoted by a combination of pcDNA3.1-VEGF(165) and gelatin sponge.

[The effect of pcDNA3.1-vEGF165 recombined vector on wound healing and the expression of collagen type I, III mRNA in wounded tissue].

OBJECTIVE: To examine the effect of pcDNA3.1-VEGF165 vector to the angiogoiesis, expression of collagen type I and type III mRNA in soft tissue injury model. METHODS: Thirty two Sprague-Daulay rats,weighted (180 +/- 20) g, were made tissue injury in the bilateral of vertebral region. Round wound (diameter 12 mm) was made by perforex on the back, removed the skin and 2 mm muscle, one side was experimental group by random and the other as control. The wound was done with sodium chloride (0.2 ml) in the control group, with the recombinant VEGF165 vector (0.2 ml, 200 mg) in the experimental group. The wound healing and other general state of health was observed after the operation. The specimens were obtained at 3,5, 7,14 and 30 days after injury. Draw the materials from the rats at the same time, all samples were divided into two parts. one ( > 0.1 g) was conserved in refrigerator at - 80 degrees C, which was extracted total RNA by TRIZOL, design the primer of rat's collagen type I and type III, RT-PCR analysis indicated that collagen type I, III. The other was fixed by 10% formalin. Examine wound healing of local tissue and count it' s MVD by HE staining. RESULTS: All the rabbits were well alive, no death or infection. Wound healing time was shorter than the control one (14.2, 17.4 d). Inflammatory cell infiltrate, cellula intersitialis, fibroblast, collagen and the density of angiogenesis were more in the experimental group than in the control one. The MVD was significant difference between the two groups at 1, 2 weeks are 63.38 +/- 9.20, 52.72 +/- 7.06 and 76.64 +/- 12.27, 66.84 +/- 9.82 (P < 0.05). The expression of collagen type I , III mRNA was found in the third day, the peak was in the second week and then degression. The collagen type I , III mRNA and beta-actin specificitic belt were found and its initial template volume different, the results was trend of RT-PCR obtained. CONCLUSIONS: The local application of pcDNA3.1-VEGF165 can enhance the expression of collagen type I, III mRNA, enhance angiogenesis and extra cellular matrix, both of which can shorten healing time of tissue injury.