Objective To investigate the risk and prediction factors for tracheotomy after cervical spinal injuries. Methods A retrospective analysis was done on 1 064 cases suffering from cervical spinal injuries, of which, according to nerve function evaluation criteria of American Spinal Injury Association (ASIA), there were 243 cases at Grade A,327 at Grade B, 306 at Grade C and 188 at Grade D. Except for seven cases with brain injuries, trachea injuries, with a tracheotomy before hospitalization, a total of 106 cases needed tracheotomy during their hospitalization. The following factors were evaluated to predict the possible causes for tracheotomy: segment of injury, ages, smoking history, past diseases (such as diabetes mellitus, hypertension and coronary heart disease) and complicated chest or lung injuries. Results The highest rate of tracheotomy for cervical spinal injury at Grade A was 35.0% (85/243), while the rate of tracheotomy for incomplete injury at Grades B, C and D was only 2.6% (21/814). Of cervical spinal injury at Grade A, all C_3 segment injuries needed tracheotomy. Of all, the percentage for C_4 and C_5 segment injuries accounted for 43.2% (105/243),of which the percentage for tracheotomy was 74% (63/85). The possibility of tracheotomy decreased gradually from below C_5. Besides C_3 segment injuries, C_4 segment injuries had the highest possibility of tracheotomy, with statistical difference compared with other segments (P

Objective patients with pSS and 30 cases of healthv contrels.The streptavidin immunohistochemical staining was performed to detect the expression and distribution of MMP-9 and CD147 in labial salivary glands.Quantitative analysis was performed by image analysis software-image plus 5.0 at the site of positive expression of MMP-9 and CD147.The correlation between their expression and the infiltrating lymphocyte foci per 4 mm2 of labial gland was analyzed by SPSS software as well as the correlation between the expression of MMP-9 and CD 147 in the salivary glands of patients with pSS.Results MMP-9 was hiKhly expressed in labial salivary glands from 52 patients with pSS and 30 healthy controls,but the expression of MMP-9 in pSS was stronger compared with that of healthy controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of MMP-9 in labial salivary glands from 52 patients with pSS(P<0.01).CD147 was highly expressed in labial salivary glands from 52 patients with pSS and 23 healthy controls，but over-expression of CD147 in PSS was more Drominent compared with that of controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of CD147 in labial salivary glands from 52 patients with pSS(P<0.01).The expression of MMP-9 and CDl47 was detected in ductal and acinar epithelial cells,lymphocyte foci in pSS.There was linear correlation between the expression of MMP-9 and CDl47 in the salivary glands of patients with pSS(P<0.01).Conclusion The results of this study suggest that the abnormal expression of MMP-9 and CD 147 is involved in the pathogenesis of pSS and play a crucial role.The interaction of MMP-9 and CD147 may be one of theimportant mechanisms leading to labial salivary glands destruction found in pSS.

Purpose To study the clinicopathological features of lymphangiomyomatosis (LAM) of pelvis lymph node.Methods A patient with endometrial endometrioid adenocarcinoma and LAM was analyzed including clinical data and pathological features.HE and immunohistoehemistry of EnVision stainings were used,and the literatures were reviewed.Results Well-moderately differentiated endometrioid adenocarcinoma could be observed in the endometrium.Proloferation of LAM cells were seen in the capsule and medulla of the pelvic lymph node.The LAM cell was spindle,epitheliod and polygonal cells with oxyphilic or clear cytoplasm which arranged surrounding lacunes.The LAM cells showed no atypia and mitosis could not seen.The tumor cells showed diffusely positive for SMA,Caldesmon,desmin,vimentin,ER and PR,the cells lining the lacunes were positive for CD34 and D2-40.The epitheliod cells were positive for HMB-45 and negative for Melan-A.The Ki-67 immunostaining showed a proliferation index of ＜ 1％.Conclusion LAM is an uncommon neoplastic multisystem disease that affects the lungs mostly.Endometrial endometrioid adenocarcinoma with LAM of pelvic lymph node is extremely rare.The diagnosis can be made according to the histological characteristics and immunohistochemical features.Moreover this conclusion will provide the clinicopathological materials for the future study about LAM.

To investigate the improving effect of inter-chain disulfide formation on protein trans-splicing, we introduce a Cys point mutation at Tyr(664) in heavy chain and at Thr(1826) in light chain of B-domain-deleted FVIII (BDD-FVIII). By co-transfection of COS-7 cell with the two Cys mutated chain genes, the intracellular protein splicing, inter-chain disulfide formation, secreted BDD-FVIII and bioactivity in culture supernatant were observed. The data showed that a strengthened spliced BDD-FVIII with an inter-chain disulfide detected by Western blotting and an elevated secretion of spliced BDD-FVIII (128 +/- 24 ng mL(-1)) compared to control (89 +/- 15 ng mL(-1)), assayed by a sandwich ELISA. A Coatest was performed to assay the secretion of bioactivity in culture supernatant and shown a much higher value (0.94 +/- 0.08 u mL(-1)) compared to that of control (0.62 +/- 0.15 u mL(-1)). It suggests that inter-chain disulfide formation could improve protein trans-splicing based dual-vector delivery of BDD-FVIII gene providing experimental evidence for ongoing in vivo study.

0.05).In the group with primary common bile duct suture,the occurrence of bile leakage was relative with hyperglycemia(P0.05).Conclusion It is the key factors,including chosing appropriate patients, intraoperative special examination, careful manipulation and effective medical treatment that can reduce the morbidity of bile leakage.

Objective:To express and purificate catalytic domain of murine?-1,3-galactosyltransferase and provide the feasible method in the tumor cell surface production ?-gal epitopes..Methods:This research established expression system in pET-15b to express catalytic domain of murine ?-1,3-galactosyltransferase,then identified its activity by HPLC with anion exchange column.Results:(1)Constructed successfully recombinant ?-1,3-galactosyltransferase catalytic domain with His-tag.(2)?-1,3-galactosyltransferase with His-tag in a soluble form was expressed and purificated effeciently.(3)Its activity by HPLC with anion exchange column showed.Conclusion:This research shows ?-1,3-galactosyltransferase in a soluble form has been expressed successfully.

Aim To investigate the effects of rosmarinic acid on H2O2-induced apoptosis in rat VSMCs and its related mechanisms. Methods Flow cytometry was used to determine apoptotic rate of VSMCs. nuclear staining by acridine orange for morphologic change,MTT assay for cell viability and Western blot for expression of Bcl-2,Bax, fas and FasL. Results After being treated by H2O2(0,250,500,750 ?mol?L-1),apoptotic rate of VSMCs was 3.71%?0.56%,17.6%?6.92%,34.9%?2.55 %, and 85.6%?5.22% by FACS analysis,and VSMCs appeared nuclear condense and nuclear fragmentation after being treated by 500 ?mol?L-1 H2O2 for 24 hours,which were typical morphological changes of apoptosis. ① VSMCs treated with 500 ?mol?L-1 H2O2 for 24 hours had a significantly decrease on cell viability compared with control,and the apoptosis rate was increased to 35.7%?1.33%; Bcl-2 protein expression decreased,Bax protein expression increased, Bcl-2/Bax protein ratio decreased and Fas receptor and Fas ligand expression also increased.② Pre-incubation with rosmarinic acid(10, 20, 40 ?mol?L-1)for 30 min enhanced the cell viability, and decreased the apoptotic rate to 31.1%?1.38%,21.2%?1.18%,13.6%?0.51% in a dose-dependent manner. Moreover, Bcl-2/Bax protein ratio increased and Fas and FasL expression decreased. Conclusions ① Rosmarinic acid antagonizes H2O2-induced apoptosis of VSMCs. ② The antagonism of rosmarinic acid on apoptosis may be correlated with an increase Bcl-2/Bax protein ratio and a decrease expression of Fas and FasL protein.

Objective To study the method and effect of lateral arm free flap in reconstruction of severe atresic eye socket.Methods Forteen cases of severe atresic eye socket,from June,2011 to June,2013,were repaired by lateral arm free flap.The flaps were designed and harvested as drop shape with size about 6 cm × 10 cm and then were removed epidermis except distal 6 cm × 6 cm area which were transferred to orbit for eye socket reconstruction.The remaining fascia and dermis were filled to augment temporal defect.Superficial temporal artery was anastomosed with posterior branch of radial collateral artery in 14 cases and superficial temporal vein was anstomosed with radial collateral vein in 11 cases,with middle temporal vein in 3 cases.Results All 14 cases lateral arm free flaps survived with no donor site morbidity.Followed up for 1 year to 3 years,artificial eye could be fitted satisfactorily and temporal contour improved.Conclusion Lateral arm free flap is a recommendable option for severe atresic eye socket reconstruction because of concealed donor site scar,proper volume,matched vascular caliber and minor donor site morbidity.

The mutation of cystic fibrosis transmembrane conductance regulator (CFTR) gene leads to an autosomal recessive genetic disorder cystic fibrosis (CF). The gene therapy for CF using adeno-associated virus (AAV) vectors delivering CFTR gene is restricted by the contents limitation of AAV vectors. In this study the split CFTR genes severed at its regulatory domain were delivered by a dual-vector system with an intein-mediated protein trans-splicing as a technique to investigate the post-translational ligation of CFTR half proteins and its function as a chloride ion channel. A pair of eukaryotic expression vectors was constructed by breaking the human CFTR cDNA before Ser712 codon and fusing with Ssp DnaB intein coding sequences. After co-transfection into baby hamster kidney (BHK) cells followed by transient expression, patch clamps were carried out to record the chloride current of whole-cell and the activity of a single channel, and the ligation of two halves of CFTR was observed by Western blotting. The results showed that the intein-fused half genes co-transfected cells displayed a high whole cell chloride current and activity of a single channel indicating the functional recovery of chloride channel, and an intact CFTR protein band was figured out by CFTR-specific antibodies indicating that intein can efficiently ligate the separately expressed half CFTR proteins. The data demonstrated that protein splicing strategy could be used as a strategy in delivering CFTR gene by two vectors, encouraging our ongoing research program on dual AAV vector system based gene transfer in gene therapy for cystic fibrosis.

We recently demonstrated that an intein-mediated protein splicing can be used to transfer B-domain-deleted FVIII (BDD-FVIII) gene by a dual-vector. In this study, we observed the effect of a variant heavy chain with six potential glycosylation sites of B domain and L303E/F309S mutations in its A1 domain, which were proven to be beneficial for FVIII secretion, on secretion of spliced BDD-FVIII. By transient co-transfection of cultured 293 cells with intein-fused variant heavy chain (DMN6HCIntN) and light chain (IntCLC) genes, the culture supernatant was analyzed quantitatively by ELISA for secreted spliced BDD-FVIII antigen and by a chromogenic assay for bioactivity. The data showed that the amount of spliced BDD-FVIII protein and coagulation activity in culture supernatant from DMN6HCIntN plus IntCLC co-transfected cells were up to (149 +/- 23) ng x mL(-1) and (1.12 +/- 0.14) u x mL(-1) respectively greater than that of intein-fused wild type heavy (HCIntN) and light chain (IntCLC) co-transfected cells [(99 +/- 14) ng x mL(-1) and (0.77 +/- 0.13) u x mL(-1)] indicating that the variant heavy chain is able to improve the secretion of spliced BDD-FVIII and activity. A cellular mechanism-independent BDD-FVIII splicing was also observed. It provided evidence for ongoing animal experiment using intein-mediated dual-AAV vector technology for delivery of the BDD-FVIII genes.