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.

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.

Objective To analyze the clinical features,diagnosis and treatment of fibromyalgiav syndrome (FMS).Methods Retrospectively analyzed 52 cases of FMS treated in our hospital between July,2008 to July,2011,and its clinical features,laboratory examination results,treatment and prognosis were analyzed.Results The ratio of female and male patients was 9∶1 in 52 FMS patients.The diagnosis was delayed for four years in average.Pain of FMS was diffuse and tenderness was symmetrical.Fatigue and sleepdisorder were the most common symptoms.There were no specific laboratory tests for this condition.Symptoms of 21 patients (57％) were alleviated among 37 patients who received drug treatment only,and comparatively,the symptoms of 12 patents (80％) were alleviated among 15 patients who received combined drugs and non-drug treatment.Conclusion The incidence of FMS is high and its clinical features vary.Clinician should pay much attention to the disease.Because the study sample size is small,so further study is needed to demonstrate the superiority of the combined therapy.

Objective To study the related factors in diabetic patients with lOwer extremital deep vein thrombosis(DVT) and the nursing strategy. Methods 58 diabetic cases underwent lower limbs 99mTc-MAA veins imaging and/or ultrasunography. The related laboratory data such as fasting plasma glucose(FPG),blood cholesterol, blood triglycerides and clinic informations such as age, disease courses, lower limbs symptoms (swelling, erooted veins and diabetic foot) and acute complication (diabetic ketoacidosis and hyperosmolar non ketotic diabetic coma) were collected simultaneously. The relationship of each factors with DVT incidence were analyzed. Results 28 patients (48.3%) complicated with lower limbs DVT. The DVT incidence was signifi-cantly different among patients with different age, disease courses, FPG, blood cholesterol, blood triglycerides, lower limbs symptoms and acute complication, respectively. No significant difference was seen in patients with different blood pressure, body weight index, diabetes type. Conclusions Various dangerous factors accumu-lating in DM such as old age, long disease courses, high FPG, high blood cholesterol and high blood triglyc-erides contribute to the high incidence of lower limbs DVT. Early intervention, comprehensive treatment and in-tensive nursing should be applied to reduce the incidence of DVT.

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.

Objective To study the related risk factors of pulmonary embolism(PE) in diabetic patients.Methods 58 diabetic cases were underwent lower limbs 99mTc-MAA veins imaging(and/or ultrasonography) and pulmonary perfusion imaging.The related laboratory data [fasting blood glucose(FBG),blood cholesterol,blood long chain triglycerides(LCT)]and clinial informations [age,disease courses,chest symptoms(chest pain and short of breathe),lower limbs symptoms(swelling,crooted veins and diabetic foot) and acute complication(diabetic ketoacidosis and hyperosmolar non ketotic diabetic coma)]were collected simultaneously.SPSS was used for χ2-test and Logistic regression analysis.Results 28 patients(48.3%) were showed to be with lower limbs deep vein thrombosis (DVT) and by 99Tcm-MAA imaging,10 cases(17.2%) with PE.The PE ratio(32.1%) of the patients with DVT was more higher than no DVT(3.3%)(χ2 = 6.53,P＜0.05).Single factor analysis showed the PE ratios had significant difference to factors of chest symptoms,lower limbs symptoms and acute complication(all P＜0.05),respectively.There were no significant difference in other factors of age,disease courses,FBG,blood cholesterol,LCT,blood pressure,weight index,DM type(all P＞0.05),respectively.Multiplicity analysis indicated:the related risk factors of PE included chest symptoms(Score = 13.316,P＜0.01) and lower limbs symptoms(Score = 7.780,P＜0.01).There were no significant differences in other factors(all P＞0.05),respectively.Conclusion The serious DM with chest symptoms,lower limbs symptoms and/or DVT could be controlled as early as possible by all kinds of treatment.It would decrease the PE complication.

Objective To evaluate body surface localization by virtue of lnnova CT reconstruction combined with DSA guidance in performing percutaneous fine needle biopsy of lung lesions. Methods Twenty-one patients with solid lung lesions were enrolled in this study. Before percutaneous lung needle biopsy, DSA rotation scanning was performed. Based on the reconstructed lnnova CT images, precise puncture scheme was formulated. Then, under DSA guidance, pereutaneous fine needle biopsy of lung lesions was carried out. Results A total of 41 percutaneous fine needle biopsies was made, of which definite pathologic diagnosis was obtained in 40, with a positive rate of 97.6%. Pneumothorax occurred in one patient (4.8%). Conclusion Percutaneous lung needle biopsy with the help of body surface localization, which is determined by Innova CT reconstruction images, and DSA guidance is a safe and technically-simple procedure with high accuracy in localization. It is of great value in the diagnosis of pulmonary space-occupying lesions.

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-tansfected 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.