Objective To explore the feasibility of nasolabial sulcus flap transfer with autologous free skin graft to repair the alar defects after malignant tumor resection.Methods From January 2012 to January 2015,9 patients with malignant tumor were treated in the hospital.After complete tumor removal,the defect area being reconstructed was 1.5 cm × 1.3 cm to 2.5 cm × 2.5 cm.The defects of 9 patients were all restored with nasolabial sulcus flap combined with autologous free skin graft.Results The 9 patients were followed up for 6-18 months postoperatively.The nasolabial sulcus flap and autologous free skin graft were survived completely in all cases.Symmetrical alae were noted with slight edema within nasal cavity but without difficult ventilation.Scar was repaired in phase-two surgery.Conclusions Nasolabial sulcus flap combined with autologous free skin graft is an optional way in alar defects restoration.Further with secondary morphologic plasty,satisfactory surgical outcome can be achieved.

Adult ; Empyema, Pleural ; complications ; etiology ; Humans ; Male ; Neck Injuries ; complications ; Tracheoesophageal Fistula ; complications ; etiology

Objective Allergic rhinitis (AR) is studied extensively while nonallergic rhinitis (NAR) insufficiently in the recent years .The aim of this study is to describe the inflammation characteristics of different types of NAR . Methods Using the skin prick test , we investigated the characteristics , nasal cytokine levels , serum cytokine levels , and the proportion of peripheral blood Treg cells in 12 cases of AR, 10 cases of NAR with eosinophilia (EOS) syndrome (NARES), 12 cases of NAR without ES (NAR), and 11 control adults . Results The NARES patients had a signifi-cantly higher level of IFN-γ(28.89 [10.97-127.07] pg/mL) than the control (8.98 [7.88-14.90] pg/mL) and the NAR patients (7.92 [7.67-45.85] pg/mL) ( P<0.05) but a lower level of nasal IL-10 than the control ([3.97±0.68] vs [4.80±1.32] pg/mL, P<0.05) .The contents of nasal IL-4, serum IL-4, nasal IL-17 and ser-um IL-17 were all markedly higher in the AR and NARES groups than in the control (P<0.05).The proportion of CD4+CD25+FOXP3+Treg cells in the CD4+T cells in the peripheral blood was (4.5±1.3)%in the AR group and (4.0±1.8) %in the NARES group, both significantly lower than (6.5±1.0) %in the control group (P<0.05) and (6.5±1.0) %in the NAR group (P<0.05). Conclusion NAR was classified according to the EOS level into NARES and NAR without EOS, which had different mechanisms and manifestations of inflammation and similar inflammatory manifestations of Th 2 and Th17.The classification of rhinitis by the level of EOS has more practical significance .

Objective To explore the influence of excessive PTEN expression to fibroblast cycle and collagen secretion induced by LPS.Methods Normal skin fibroblast in the patient with hyperplastic scar were cultured in vitro.When the primary culture was close to 80% fusion,the digestive passage was performed,cultured to the third generation.LPS(0.5 μg/mL) was adopted to stimulate the third generation of normal skin fibroblasts.Defective adenovirus carrying PTEN gene was transfected to the third passage fibroblasts after LPS stimulation.Flow cytometer was adopted to detect the cell cycle.ELISA method was adopted to detect the secreted collagen amont.Results Excessive PTEN expression could inhibit the increase of G2M cell cycle induced by LPS.LPS stimulation could increase the secretion of collagen in skin fibroblasts,yet excessive PTEN expression could inhibit the secretion of collagen induced by LPS.Conclusion LPS could increase the amont of fibroblasts on G2M cell cycle and secretion of collagen,yet excessive PTEN expression can inhibit the effect.

Objective To study the experimental results of selective reinnervation of posterior cricoarytenoid(PCA) muscles by upper root of unilateral phrenic nerve.Methods Eight beagle dogs were used in this experiment.The left recurrent laryngeal nerves (RLNs) were severed and anastomosed with upper root of phrenic nerve, and the intralaryngeal adductor branch was cut and sutured into the belly of the ipsilateral PCA muscle. The right RLNs were kept intact, and were used as control group.Videolaryngoscopy and electromyography (EMG) were performed at preoperative, immediately after surgery and 6 months after surgery.After completion of all physiologic testings,the dogs were sacrificed and bilateral PCA muscles and intralaryngeal part of recurrent laryngeal nerves were harvested, then histological examination was carried out.The laryngeal nerve was stained with toluidine blue and the morphology of the axons was observed under light microscope.Results Preoperatively, the movement of bilateral vocal folds were normal in all dogs.The left vocal folds were fixed immediately after surgery, 6 months after surgery, the left vocal folds in all 8 dogs recovered inspiratory abductive movement.Spontaneous and evoked electrical activities of the reinnervated PCA muscles could be recorded in all cases during inspiration preoperatively.The left PCA muscles were electrical silent during normal inspiration, and evoked activities were not induced immediately after surgery.Spontaneous electrical activities and evoked electrical activities were recorded 6 months after surgery, and had no significant difference when compared with those of postoperative (P>0.05).There was no significant difference in the cross-sectional area of the bilateral posterior cricoarytenoid muscles fibers after the masson staining.Muscle collagen relative cross-sectional area and collagen relative cross-sectional area and muscle/collagen cross-sectional area ratio difference were not statistically significant(P>0.05).Nerve pulp numbers of left and right sides of the recurrent laryngeal nerves had no significant difference (P>0.05).Conclusion Selective reinnervation of posterior cricoarytenoid muscles by the upper root of unilateral phrenic nerve can restore inspiratory vocal fold abduction to a satisfactory extent and avoid aberrant regeneration.

BACKGROUND: The activation of hypothalamus-pituitary-adrenal cortex axis may play key role in the increasing expression of hypothalamic corticotropin-re-leasing hormone (CRH) during stress reaction. However by what way to induce the CRH expression in hypothalamic neuron, and whether CRH can activate hypothalamic neurons are still not very clear.OBJECTIVE: To observe the changes of intracellular cyclic adenosine monophosphate (cAMP) and cytoplasmic Ca2+ concentration in the hypothalamic neurons cultured in vitro due to exogenous CRH stimulation.DESIGN: Comparative observation experiment.SETTING: Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA; Department of Neurosurgery , Tiantan Hospital, Capital University of Medical Sciences MATERIALS: This experiment was carried out in the Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA between December 1999 and March 2002. Hypothalamus was obtained from fetus rat at pregnancy of 17 days for the in vitro culture of hypothalamic neurons.METHODS: Hypothalamic neurons were co-cultured with exogenous CRH,with or without pretreatment with specific CRH 1 receptor antagonist -CP-154526. hypothalamic neurons were randomized into: ① CRH (10-12,10-10, 10-8, 10-6 mol/L) stimulation group. ② CP-154526(500 μmol/L)pretreatment aud CRH ( 10-12, 10-10, 10-8,10-6 mol/L) stimulation group. ③Hypothalamic neurons in corresponding normal control group were exposed to the isotonic saline stimulation. PTI fluorescence image system was used to determine and analyze the change of cytoplasmic free Ca2+ concentration in hypothalamic neurons due to exogenous CRH stimulation and RIA was used to detect the neuronal cAMP content.MAIN OUTCOME MEASURES: ①Cytoplasmic free Ca2+ concentration in hypothalamic neurons. ②cAMP content in hypothalamic neurons.RESULTS: The cytoplasmic free Ca2+ concentration and cAMP content were relatively lower in the hypothalamic neurons in normal control group,which obviously increased due to CRH stimulation [(240±22),(153±11)nmol/L; (3.26±0.19),(0.44±0.02) pmol/dish,P ＜ 0.01];CP-154526 could remarkably suppress the CRH (10-6 mol/L)induced increase in cytoplasmic free Ca2+ concentration and cAMP content in hypothalamic neurons [Ca2+ concentration: (240±22),(171±16)nmol/L; cAMP content:(3.26±0.19), (2.33±0.21) pmol/dish, P ＜ 0.01].CONCLUSION: CRH can directly act on hypothalamic neurons via type 1-receptor,thereby increase the cytoplasmic free Ca2+ concentration and cAMP content in hypothalamic neurons,playing the key role in the modulation of the synthesis and secretion of CRH during the activation of hypothalamic neurons.

BACKGROUND: Through what signal pathway does corticotropin release hormone (CRH) regulate hypothalamic neuronal neuroendocrine activity during acute stress?OBJECTIVE: To probe into the regulatory effects of CRH on CREB secretion in hypothalamic neurons.DESIGN: Repetitive measurement design.SETTING: At Field Surgery Research Institute of Daping Hospital, Third Military Medical University of Chinese PLA; Neurosurgery Department,Tiantan Hospital Affiliated to the Capital University of Medical Sciences.MATERIALS: This experiment was carried out in Daping Hospital of Third Military Medical University of Chinese PLA between December 1999and March 2002. Rat fetuses were selected from Wister rats of 17-day gestation.METHODS: In vitro cultured cells were divided into the following groups:① CRH (10-12, 10-10, 10-8 and 10-6 mol/L) stimulation groups. ② Pretreated with nimodipine (5 μmol/L) or CP-154526 (500 μmol/L) followed by CRH (10-12, 10-10, 10-8 and 10-6 mol/L ) stimulation groups. ③ Corresponding control groups stimulated with isotonic physiological saline. PTI fluorescence imaging system was used to detect the changes of neuronal cytoplasmic free calcium concentration; meanwhile, Western blot technique was used to determine the changes of neuronal P-CREB content.MAIN OUTCOME MEASURES: ① Changes of neuronal cytoplasmic free calcium concentration. ② Changes of neuronal P-CREB content.RESULTS: The content of cytoplasmic free calcium in hypothalamic neurons was lower in normal control group, and it increased immediately after exogenous CRH stimulation. However, such increase could be suppressed by pretreatment with nimodipine or CP-154526 before CRH stimulation,and the increase of neuronal P-CREB content was also obviously suppressed.CONCLUSION: During acute stress, the combination of CRH with hypothalamic neuronal CRH 1 receptor leads to the opening of membrane Ltype calcium ions channels, thus enhancing the influx of calcium ions and increasing cytoplasmic free calcium ions content, which would further activate P-CREB signal transduction pathway in neurons. It suggests that CRH may play a vital role in hypothalamic neuronal activation.

Objective To construct a gene recombinant lentiviral vector pCMV -G -U6 -hHGF and detect its expression in C2C12 myoblast cells .Methods hHGF gene fragments were obtained and purified by RT -PCR method ,and were cloned to pCMV -G&NR -U6 ,then the restructured lentiviral vector was transformed into e . coli DH5 alpha ,the positive colonies were identified by BamHI and Hind Ⅲ enzyme digestion .The selected positive colonies were tested by PCR and sequencing analysis .The expression plasmids and packing plasmids were co -trans_fected into 293 T cells ,and virus titer was observed under the fluorescence microscope .Furthermore ,transfected C2C12 cells with lenti virus ,and the expression of HGF was detected by PCR and WB methods .ResuIts PCR and sequencing analysis showed that the lentiviral vector was constructed correctly and successfully ,the virus titer was above 1 x 109 IU/mL .The results of PCR and WB showed that HGF expression level in the lentiviral vector group was much higher than those of in blank control and negative control groups ,and yet the expression was stable after 72 hours .ConcIusion The lentiviral vector pCMV -G﹠NR -U6 -hHGF has been successfully constructed ,and stable expressed in C2C12 cells .It provides references for experimental study in the fields of the denervated skeletal muscle fibrosis and nerve regeneration treatment .

Objective To study the curative effects and characteristics of vocal cord shallow lamina propria resection on the treatment of vocal cord leukoplakia .Methods A total of cases of vocal cord leukoplakia were re-ceived vocal cord mucosa stripping surgery (69 cases) and vocal superficial lamina propria resection with or without suture (69 cases) respectively during January 2006 - December 2011 ,and all cases were taken dynamic laryngosco-py and voice acoustic analysis before surgery ,at 2 weeks ,4 weeks ,6 weeks ,8 weeks ,3 months ,6 months ,and 12 months after surgery .We observed the curative effects and characteristics after operation of two different surgery on the treatment of vocal cord leukoplakia with precancerous lesions .Results Two week after operation ,the vocal cords mucous wave ,vocal cords vibration symmetry ,regularity ,total hoarseness degree (G) ,Jitter ,Shimmer , NHRvaluesinthe2groupsweresignificantlylowerthanthoseofpreoperation(P<0.05),buttherewerenosignif-icant differences between the two groups(P>0 .05) .The three main index of dynamic laryngoscope ,voice acoustic parameters at 4 weeks after operation were significantly lower than those at 2 weeks after operation in the vocal cord mucosa stripping surgery group ,the difference were statistically significant (P<0 .05) ,but it showed no statistical difference between the group at 6 weeks after operation and the groups at other time points (P>0 .05) .The three main index of dynamic laryngoscope ,voice acoustic parameters at 6 weeks after operation were significantly lower than those at 2 ,4 weeks after operation in the vocal cord shallow lamina propria resection group ,the difference were statistically significant (P<0 .05) ,but it showed no statistical difference between the group at 8 weeks after operation and the groups of other time points (P>0 .05) .The voice restoration was faster in the vocal cord mucosa stripping surgery group .The recurrence rate was lower in the vocal cord shallow lamina propria resection group than those in the traditional vocal cord mucosa stripping surgery group ,the difference was statistically significant (P<0 .05) . Conclusion The vocal cord shallow lamina propria resection is a minimally invasive operation for the treatment of vocal cord leukoplakia ,with low recurrence rate and good the voice recovery .

Objective Nonallergic rhinitis with eosinophilia syndrome (NARES) is associated with such diseases as bronchial asthma , nasal polyps , and aspirin intolerance , but there is a lack of studies on its inflammatory conditions .The aim of this study is to describe the inflammation characteristics of NARES . Methods This study included 101 cases of allergic rhinitis ( AR) , 39 cases of NARES, and 162 adult controls .We analyzed the inflammation char-acteristics of the patients using skin prick test ( SPT) , nasal douche , fractional exhaled nitric oxide ( FeNO ) measurement , serum eosino-phil (EOS) counting, serum total IgE (tIgE) determination, induced sputum detection, and nasal and bronchial provocation tests . Results Compared with the controls , the NARES and AR groups showed significant increases in the positive rate of nasal provocation (32.1%vs 69.2%and 75.2%, P<0.05), positive rate of bronchial provocation (1.2%vs 10.3%and 14.9%, P<0.05), and FeNO level ([15.70 ±5.20] ppb vs [37.25 ±22.95] and [39.00 ±24.29] ppb, P<0.05), as well as in the serum EOS and tIgE levels, EOS count, and ratio of EOS in the induced sputum (P<0.05).However, the level of serum tIgE was significantly lower in the NARES than in the AR group (53.3 [23.3-186.0] kU/L vs 197.0 [62.6-391.0] kU/L, P<0.05). Conclusion NARES is a syndrome with nasal, lower airway, and systematic inflammation, similar to AR in inflammatory intensity.Therefore, for patients with NARES, attention should be paid not only to upper airway but also to lower airway and systemic inflammation .