Objective To investigate the effects of DMOG on the microcirculation of the choke-area and the survival of the cross-boundary flap in rats via tail vein injection.Methods Rats with ischemic three-territory perforator flaps on the dorsum were treated with DMOG at a dosage of 40 mg/kg body weight via tail vein injection at 1 day before surgery(day-1),the time of surgery(day 0),1 day after surgery(day 1),2 days after surgery(day 2) and 3 days after surgery(day 3).Control group received sterile saline at the same time points and same dosage via tail vein injection.① Draw materials from the choke-area at day 1,day 3 and day 7,HE stain was used to compare the diameter size of the artery and vein at the same site.② Western blotting to check the expression of PCNA and HIF-1α,ELISA to detect the content of PCNA,HIF-1α,SDF-1α and VEGF at day 7.③At day 7,measure the survival area of the flap and observe the vessel of the flap by lead oxide-gelatine technique.Results ① There was a greater survival rate of (96.3 ± 5.1)％ in the treatment group than in the control group with (73.9 ± 5.8)％ at day 7 (P ＜ 0.05).② The diameter size of the arterioles and venules were dilated in both groups until postoperative days 7.But the treatment group was more expanded than the control group at day 3(2.20 ± 0.26 vs.1.50 ± 0.20,P ＜ 0.05) and day 7(3.67 ± 0.35 vs.2.03 ± 0.15,P ＜ 0.05).③ The skin expression of PCNA and HIF-1α in the treatment group were greater than the control group(P ＜ 0.05) at day 7.④ The content of skin PCNA in the treatment group and control group were(8.95 ± 0.71) ng/mg and (4.15 ± 0.72) ng/mg,HIF-1α were(5.04 ± 0.50)ng/mg and (2.98 ± 0.29) ng/mg,SDF-1α were (2.91 ± 0.61) ng/mg and (1.39 ± 0.62) ng/mg,and VEGF were(2.17 ± 0.41) ng/mg and (0.95 ± 0.44) ng/mg,respectively.The treatment group was greater than the control group (P ＜ 0.05).Conclusion DMOG can improve the microcirculation of the choke area,and then increase the survival of the perforator skin flaps in rats via tail vein injection.

Objective To investigate the effects of human amniotic membrane on the survival of the crossboundary perforator flap in rats through distal subcutaneous implantation.Methods From February,2016 to December,2016,38 SD rats weighing 270-300 g were randomly divided into experimental group and control group (n=19).A three-territory perforator flap (3 cm× 10 cm) with the iliac artery as the pedicel elevated on the right dorsum of the rats.Experiment group,a section of amnion was sutured to the distal subcutaneous of the flap and then the flap was sutured back to its donor site.Control group,after the flap elevated,directly sutured back to its donor site.The blood flow of the pedicel were detected by laser Doppler flowmetry at the day before surgery,1 day after surgery,3 and 7 days after surgery,with 3 rats in every group.When the rats were anesthetized,biopsies were taken from the choke area Ⅱ at the day before surgery,3 and 7 days after surgery,with 9 rats in every group.When the rats were anesthetized,HE was used to compare the diameter size of the artery and vein at the same site.At day 7,measure the survival area of the flap,and 3 of them observe the vessel of the flap by lead oxide-gelatine technique.Results The survival rate of the experimental group and the control group after 7 days were (89.09±4.23)％ and (74.56-±5.59)％ respectively,the experimental group was significantly higher than that of the control group (t=5.48,P=0.00).X-ray showed that 7 days after operation,the pedicel of iliac artery in the experimental group was bigger than that in the control group.The blood flow detection showed that the blood flow of the experimental group was higher than that of the control group (t=2.39,3.06;P=0.03,0.00,respectively) at day 1 and day 3 after surgery.Hematoxylin eosin staining showed that arteriovenous tube diameter at the Choke Ⅱ were gradually increased on the day3 and day7 after surgery in both group,but the experimental group diameter expanded multiple was higher than that of control group (t=3.52,3.50;P=0.02,0.02,respectively).Conclusion Human Amnion subcutaneous embedding may improve the blood flow of the vascular pedicle,expand the microvascular at Choke area Ⅱ,improve microcirculation,as a result to promote the survival of cross perforator flap,while the mechanism is needed to understanding.

Objective: To observe the clinicopathologic features of oropharyngeal squamous cell carcinoma associated with human papilloma virus (OPSCC-HPV) and discuss the role and value of different in situ hybridization (ISH) detection methods for HPV in pathologic diagnosis. Methods: Fifteen cases of OPSCC-HPV were collected from Department of Pathology, Beijing Tongren Hospital, Capital Medical University from January 2016 to August 2018. These cases were diagnosed in accordance with the WHO classification of head and neck tumors. The histopathologic features and the clinicopathologic data were retrospectively analyzed. Immunohistochemistry (two-step EnVision method) was done to evaluate the expression of p16, Ki-67 and p53. ISH was used to detect HPV DNA (6/11 and 16/18). RNAscope technology was used to evaluate the presence of HPV mRNAs (16 and 18). Results: The mean age for the 15 patients (8 males, 7 females) was 47 years (range from 30 to 69 years). OPSCC-HPV typically presentedat an advanced clinical stage, six patients had cervical lymphadenopathy (large and cystic), seven had tonsillar swelling, one had tumor at base of tongue, and one had odynophagia. Microscopically the tumors exhibited distinctive non-keratinizing squamous cell carcinoma morphology. Cervical nodal metastases were large and cystic, with thickening of lymph node capsules. OPSCC-HPV raised from crypt epithelium and extended beneath the tonsillar surface epithelial lining as nests and lobules, often with central necrosis. Tumor cells displayed a high N: C ratio, and high mitotic and apoptotic rates. Tumor nests are often embedded within lymphoid stroma, and may be infiltrated by lymphoid cells.Fifteen cases (15/15) were strongly positive for p16; Ki-67 index were 60%-90%; they were focally positive or negative for p53. Ten cases (10/10) were negative for HPV 6/11 DNA, and one case(1/10) was focally positive for HPV16/18 DNA. Eleven cases (11/11) were strongly positive for HPV16 mRNA, one case was focally positive for HPV18 mRNA. Conclusions OPSCC-HPV is a pathologically and clinically distinct form of head and neck squamous cell carcinoma. OPSCC-HPV is associated with high-risk HPV (type 16) in all cases. Detection of high-risk HPV16 mRNA by RNAscope is of great significance in the final diagnosis and pathogen identification.