Objective The aim of this study was to establish a rat model of blood hypercoagulable state by intra?venous injection of thrombin and to provide a model for researches on hypercoagulable state. Methods Rats were divided into six groups and were injected with normal saline and 2?5, 5, 10, 20, 40 U/kg thrombin solution through the femoral vein, respectively. Then, blood was drawn to test the activated partial thromboplastin time (APTT), prothrombin time ( PT) and fibrinogen ( FIB) , and to observe the death rate of rats in these groups to verify the optimal dosage. On this ba?sis, rats were injected thrombin of the best dose through the femoral vein, and blood samples were collected at 0, 10, 30, 60, 120, 180, 300 (s) to test APTT and PT and FIB for determining the best time for blood sampling. At last, the rats were divided into control group and thrombin group to inject normal saline or thrombin solution in the best dose via the fem?oral vein, and blood was taken at the best time to test APTT, PT, FIB and whole blood viscosity. Results APTT and PT values of the 10 U/kg thrombin group were the shortest, and FIB value of this group was the highest among these groups. APTT and PT values of blood sample collected at about 60 s after thrombin injection were the shortest, and FIB value was the highest. Compared with the control group, PT and APTT values of the thrombin group were shorter (P<0?05), and blood viscosity and FIB were higher ( P<0?05 ) . Conclusions Injecting thrombin solution into the femoral vein can be used to establish a rat model of hypercoagulable state. The best dose of thrombin solution is 10 U/kg in a concentration of 2 U/mL. The best time to collect blood sample is 60 s.

Objective To explore the formation mechanism of peritumoral brain edema(PTBE)by vascular endothelial growth factor(VEGF).Methods 40 biopsies were obtained from 37 patients.Inmunohistochemical staining and Western blot were performed to detect the expression of VEGF protein.Reverse-transcriptase polymerase chain reaction(RT-PCR)was used to analyze the presence and quantity of VEGF mRNA.The extent of PTBE was estimated as an edema index(EI)based on preoperative magnetic resonance imaging.Results In VEGF-positive cases,a decreasing gradient of VEGF protein expression was observed with increasing distance from tumors(0.38±0.08,0.20±0.03,0.04±0.02).In meningiomas,the protein level and the mRNA level were congruent and the expression of both protein and mRNA had a significant correlation with EI(protein: r =0.892,RNA: r =0.875,P ＜ 0.05).However,in peritumoral areas,protein level were not consistent with the mRNA level.Protein results showed high correlation with EI(r =0.912,P ＜ 0.05),but mRNA almost was almost undetectable(0.06±0.02).Conclusion VEGF is impartant on PTBE.It is concluded that VEGF macromolecules are secreted by tumor tissue and enter peritumoral normal brain tissue to induce edemagenesis in meningiomas.

Objective To investigate the role of BMSCg on enhancing the implant survival and bacheal epithelium regeneration. Methods After transplanted with cryopreserved 2 weeks and 6 weeks allocraft, PHK-26 labeled 3-5 passage BMSCs were injected into the recipient rats via tail vein. Rats in the control groups were injected with the same amount of PBS.We observed the histology of the transplanted trachea including epithelium growth and regeneration, and the PKH-26 fluorescence levels at the para-anastomotic trachea to evaluate the role of BMSC transplantation on the epithelium regeneration. Results Rats from BMSCs injection group survived a long period. Histological observation showed that the tracheal lumen was covered by psudo-striated ciliated columnar epithelium. The cartilage structure was intact. There are no signs of denaturation and necrosis. In the PBS injection group, epithelium regeneration is better in PBS-6-week group than PBS-2-week group. The longest survival time in PBS-6-week group was 32 days, whereas it was 10 days in PBS-2-week group. In BMSCs injection group, rats in BMSC-6-week groups survived longer than 8-week group(12 rats were terminated at 1 week, 4 weeks and 8weeks as planned). There was one rat who survived and were terminated at the designated 8 weeks time point (there were 8regenerated epithelium was similar in the two BMSC transplanted groups. PKH-26 labeled BMSCs migrated to the implant site and showed red fluorescence, with most red fluorescence shown at the anastomotic part. Conclusion BMSCs can migrate to the impaired tissue to repair it. BMSCs may exert their reparation function via enhancing epithelium regeneration.

Objective To observe the effect of electroacupuncture on back three-acupoints ( namely bilateral Dazhu, Fengmen, Feishu) on the protein and mRNA expression of transforming growth factor-beta 1 (TGF-β1) and Smad3 in asthmatic rat airway remodeling model, and to explore its therapeutic efficacy and molecular mechanism. Methods Rat asthma model was established by inhalation of ovalbumin. After sensitization for 6 weeks, rats were killed. And then the airway morphological parameters of rats were measured by image analysis. The protein and mRNA expression of TGF-β1 and Smad3 in lung tissues were detected by immunohistochemistry and real-time quantitative polymerase chain reaction (qRT-PCR) respectively. Results Compared with the blank group, the standardized values of muscle cross-sectional area including airway smooth muscle area (WAm) /perimeter of the basement membrane (Pbm), and bronchial inner wall area (WAi)/Pbm were increased in the model group. The protein and mRNA expression levels of TGF-β1 and Smad3 were also increased in the model group. In electroacupuncture group, the above observation indexes were decreased, and the differences were statistically significant ( P<0.05 compared with the model group). Conclusion The experimental results indicated that back three-acupoint electroacupuncture has an effect on remodeling airway, and one of the mechanisms is probably associated with the regulation of TGF-β1/Smad3 signaling pathway.

Objective To reveal the difference between the standards for genu varum and genu valgum in the medical standard directory PLA Air Force( PLAAF) for recruitment of flying cadets and those adopted in the United States Air Force ( USAF) , and suggest a method for the reform of our flying cadets recruitment.Methods The rejection rate and comprehe nsive assessment qualification rate of genu varum and genu valgum during physical examinations for recruitment of flying cadets between 2012 and 2015 were analyzed.The different standards for genu varum and genu valgum in PLAAF and USAF flying cadets recruitment were compared and subjected to an empirical study.Results During the final physical examination for selection of flying cadets between 2012 and 2015, only 18 candidates were eliminated because of genu varum and genu valgum, accounting 9.1% of the total eliminated candidates because of orthopadics diseases.Four candidates with genu varum and genu valgum passed the comprehensive assessment in 2014 and 31 in 2015, which accounted for 15%of the candidates with orthopaedics diseases approved by comprehensive assessment.The standards for genu varum and genu valgum in PLAAF were based on morphology while those in the USAF based on the function of knees.According to the USAF medical standard directory, 9 of the candidates rejected because of genu varum and genu valgum were qualified and 9 disqualified.Among the candidates with genu varum and genu valgum approved by comprehensive assessment, 32 were qualified and 3 disqualified.Conclusion The standards for genu varum and genu valgum in PLAAF medical standard directory are of lower accuracy.The standards of USAF should be referred to and the function of knees should be considered in selection of flying cadets.Femur-tibia angle should be measured to improve the morphological standards.

Objective To compraison the of exposure in the endonasal transsphenoidal approach to the sellar between microscope and endoscope. Methods Ten formalin-fixed, silicone-injected adult cadveric heads were studied. A direct endonasal transsphenoidal approach was performed via the right nostril, pushing aside the nasal septum, then reach the sphenoidal sinus. The approach was performed with the operating microscope first, then with the endoscope. For each step (sellar, suprasellar, parasellar and clival), the operative region afforded by direct microscopic view was measured and then compared with that obtained by using the edndoscope. Results It was found that the endoscope provided greater view than microscope in this approach. Although the microscope provides an adequate view of the midline structures and part of the contralateral parasellar areas; under direct endoscopic vision, the lateral extension could be widened by an additional 6.5 mm on the ipsilateral and 4 mm on the contralateral side. At suprasellar region, the microscope provides could expose the posterior part of, optic nerve and optic chiasma; but could not expose the areas anterior and superior the interspace superior the optic chiasma. Compare with the microscope, the endoscope allowed extension of bone removal and dual opening for an additional 4 mm anteriorly at the sagittal axis and an additional 3.5 mm on the ipsilateral and 4 mm on the contralateral side. At the clivus region, the medial surface of the vertical segment of the ICA and the basilar artery could be partially 7 exposed by the microscope. By the endoscope, it could gain an additional 4 mm on the ipsilateral side and 2.5 mm on the contralateral side in width. Because of the anatomical boundaries of the sphenoid sinus, the anatomincal exposure by the microscope same as the endocope at the sagittal axis. Conclusion The endoscope allows for a panoramic view and permits widening of the operative exposure in all directions. The endoscope is more suitable in the the minimal and expanded endonasal transsphenoial approach.

Objective To construct the eukaryotic expression vector of cyclin-dependent kinase ( CDK) 7 labeled with Myc tag, obtain the expressed product , and identify its interaction with FLAG-P53 at the protein level .Methods Human CDK7 coding gene region amplified from the mammary cDNA library by PCR was inserted into the pXJ -40 vector.The recombinant plasmid Myc-CDK7 transfected into human 293T cell lines was investigated and examined by SDS-PAGE and Western blotting.In addition,assay was applied to determine the interaction between Myc-CDK7 and FLAG-p53.Results The coding region of CDK7 was successfully amplified by PCR and cloned into pXJ-40 vector, which was identified by double enzyme digestion and gene sequencing .Myc-CDK7 was successfully expressed in human 293T cell lines according to SDS-PAGE and Western blotting assay indicated that Myc-CDK7 could interact with P53 protein, which verified its known function .Conclusion The eukaryotic expression vector Myc-CDK7 is successfully obtained , which will contribute to further research on CDK 7-mediated cell cycle regulation .

Objective To construct the prokaryotic expression vector of human autophagy-related LC3B gene,obtain the GST-LC3B recombinant plasmid , purify the GST-LC3B fusion protein and identify its activity in vitro.Methods Human LC3B coding region was amplified from the human mammary gland cDNA by PCR and inserted into the prokaryotic expres -sion vector pGEX-KG.The recombinant plasmid pGEX-KG-LC3B was transformed into E.coli Rossate.The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis .The function of the purified protein GST-LC3B was detected by GST pull-down assay.Results About 400 bp of the LC3B coding region was successfully amplified from the mammary gland library by PCR and inserted into pGEX -KG.The result of double diges-tion and sequencing showed that the GST-LC3B recombinant plasmid was successfully obtained .The GST-LC3B fusion pro-tein of about 40 000 (Mr) was successfully purified and identified by SDS-PAGE and Western blotting analysis.GST pull-down assay showed that GST-LC3B could interact with Atg4B, which identified its known function .Conclusion The pro-karyotic expression vector of GST-LC3B is constructed successfully , which will facilitate further research on the function of LC3B in autophagy.

Objective:To explore the expression of abnormal spindle-like microcephaly-associated (ASPM) in liver cancer tissues and clarify its prognostic relationship with clinicopathological features of liver cancer after liver transplantation.Methods:Immunohistochemistry was employed for detecting the expression of ASPM in 72 liver cancer tissues and 36 adjacent tissues of liver cancer liver transplant recipients fulfilling the Hangzhou criterion. In conjunctions with clinicopathological data, the correlation between the expression level of ASPM in liver cancer tissues and the clinicopathological characteristics and the post-transplantation prognosis for liver cancer were statistically analyzed.Results:During a median follow-up period of 29 months, 20 patients relapsed and 8 died after transplantation. Immunohistochemical results indicated that the high-expression rates of ASPM were 58.3% and 25.0% in liver cancer and adjacent tissues ( P=0.001). The difference was statistically significant. The high-expression rate of ASPM was significantly higher in liver cancer tissues than that in adjacent tissues. The expression level of ASPM was not correlated with gender, age, smoking/alcoholic history, hepatitis history, preoperative level of alpha-fetoprotein (AFP), tumor size, tumor load or vascular tumor thrombus ( P>0.05). And the postoperative high-expression rates of ASPM were 51.0% and 76.2% in pathological differentiation type Ⅰ-Ⅱ and Ⅲ-Ⅳ groups ( P=0.049). The difference was statistically significant. The wrose pathological differentiation type of liver cancer, the higher expression level of ASPM in liver cancer tissue. In liver cancer tissues, the overall 1/3/5-year survival rates of ASPM high/low-expression group were 97.6%, 80.6%, 80.6% and 93.3%, 89.7% and 89.7% respectively ( P>0.05). There was no statistical significance. And 1/3/5-year long-term disease-free survival rates were 78.6%, 55.5%, 55.5% and 86.3%, 86.3% and 86.3% respectively ( P=0.036). The difference was statistically significant. The disease-free survival rate was lower in ASPM high-expression group and post-transplantation prognosis was worse. Conclusions:The expression of ASPM is significantly higher in liver cancer tissues than that in adjacent tissues. And the expression level of ASPM in liver cancer tissues is correlated with pathological differentiation types of liver cancer and has an impact on tumor-free survival of patients after liver transplantation for liver cancer.

Objective To explore the characteristics of opportunistic infections(OIs)in HIV-infected indi-viduals with suboptimal immune reconstitution after ART treatment so as to provide a reference for preventing and managing HIV infections.Methods The clinical data including opportunistic infections specifically were acquired from 112 HIV-infected individuals with suboptimal immune reconstitution from the outpatient department of Wuming Hospital,Guangxi Medical University.The impact of baseline CD4+T lymphocyte counts on the incidence,type,and mixed infection rates of the opportunistic infections were analyzed.Results The opportunistic infection rate among the 112 HIV-infected individuals with suboptimal immune reconstitution was 42.86%,among which fungal infections were the most commonly seen.The opportunistic infection rate of the patients with a baseline of CD4+T lymphocyte counts≤50/μL was significantly higher than that of the patients with a baseline of CD4+T lymphocyte counts>50/μL,and there was no significant difference in the type of opportunistic infections as well as the rate of mixed infections.Conclusion HIV-infected people with suboptimal immune reconstitution in Guangxi are susceptible to HIV OIs.Among them,the group with a baseline CD4+ T lymphocyte counts≤50/μL has a higher rate of OIs,mainly fungal infections.