Objective To realize the construction and expression of human dsFv genes to rabies virus. Methods The genes of dsFv VH and VL were amplified by PCR-based point mutagenesis strategy. Then the genes were cloned into plasmid pET-22b (+). E. coli BL21(DE3) was transformed with the recombinant expression plasmid and the protein was induced in Luris-Bertani medium by addition of IPTG. The inclusion body proteins of VH and VL were denaturalized by GuHCl, and then re-naturalized in refolding solution to form dsFv fragments. Afterwards, we evaluated the antigen-binding activity of the dsFv and its stability as compared with its originative scFv. Results The fragment genes of dsFv to rabies virus were constructed successfully by PCR-based point mutagenesis strategy. Sequence analysis proved that cysteines were introduced into the position 44 aa of VH and position 100 aa of VL. The dsFv VH and VL genes were expressed in PET22b(+)/BL21(DE3). The VH and VL protein folded into the active dsFv antibody fragments which showed specific binding capability to rabies virus. Conclusion We succeeded in achieving the stabilization of the human scFv to rabies virus and obtaining the active human dsFv antibody fragments. This established a solid basis for further study of the biological activaty and clinical application of dsFv to rabies virus.

Objective To evaluate the clinical effects of endobronchial ultrasound - transbronchial needle aspiration (EBUS-TBNA) in the evaluation of staging of lung cancer.Methods Between July 2008 to March 2010,the first 128 patients selected by CT or PET/CT scanning with lung cancer in whom metastatic carcinoma in the hilar and/or mediastinal lymph nodes underwent EBUS-TBNA and were clinically followed up.There were 102 males and 26 femals with the age of 37 - 85 years,average 60.1 years.Review the performance in check and the result of biopsy.Results From 128 patients of mean age 60.1 years ( range 37 - 85 ),189 lymph nodes were punctured.The mean diameter of the nodes was 12.3 mm and the range was 6-16 mm.There were no procedural complications.Accuracy,sensitivity,and specificity for EBUS-TBNA were 98.53％,98.50％,and 100％,respectively.Conclusion EBUS-TBNA allows real-time visualization of mediastinal and hilar lymph nodes,allowing sampling safely and efficiently.It has great potential for diagnosis of staging of lung cancer.

BACKGROUND: Magnetic resonance imaging(MRI)is considered safe for diagnosis of meniscus injury,having accuracy as high as 80% to 100%.However,it is found that there are some false positive and false negative in the clinic.OBJECTIVE: Through assessment of menisci injuries of the knee using magnetic resonance imaging and arthroscopy,to investigate MRI manifestation in different types of meniscal injuries.METHODS: A study from January 2003 to December 2008 was performed in 212 patients with knee injury undergoing preoperative MRI.Radiograph was read by physicians who had a senior professional title from radiology and orthopaedics departments,and meniscus damage and damage morphology were predicted.Arthroscopy was conducted by chief physician.Meniscus damage during surgery was considered as gold standards.The accuracy of them was compared.The statistics of accuracy of MRI diagnosis was gathered.RESULTS AND CONCLUSION: Totally 212 patients and 230 knees were included,and a total of 215 knees of meniscus injury were described by MRI diagnosis and 213 knees of meniscus injury were proven by arthroscopy.The sensitivity,specificity and accuracy of MRI diagnosis of meniscus injury respectively were 93.5%,64.7%,93.0%.It is found that the sensitivity of MRI diagnosis of bucket-handle damage is high.MRI is easy to miss diagnosis in a light meniscus posterior horn tear,and resuts are more false positive in lateral meniscus injury.It needs arthroscope operation or further surgery in some condition such as gradeⅢ type injury,meniscus displacement after injury and articular capsule separation,which indicates instable meniscus.

BACKGROUND:The relationship between the occurrence time and clinical symptoms of bone marrow edema in nontraumatic femoral head necrosis is stil controversial. OBJECTIVE:To evaluate the relationship of bone marrow edema in nontraumatic femoral head necrosis with Association Research Circulation Osseous (ARCO) staging and pain grading. METHODS: Forty-eight patients with nontraumatic femoral head necrosis (58 hips) were confirmed to have bone marrow edema on MRI T1 weighted images and T2 fat suppression images. Then, the patients were graded according to the MRI Classification Method reported by Zhao Pei-rong, the ARCO Staging System and Harris hip score for pain grading. We analyzed the correlation of bone marrow edema with ARCO staging and pain grading. RESULTS AND CONCLUSION:The incidence rate of bone marrow edema in ARCO I-III was increased in sequence, and the difference was statisticaly significant (χ2=27.69,P=0.001); the average rank of bone marrow edema in ARCO I-III was increased significantly (χ2=19.947,P=0.000). The incidence rate of bone marrow edema also increased among different pain grading, and the difference was statisticaly significant (χ2=57.0, P=0.000); the average rank of bone marrow edema among different pain grading was increased significantly (χ2=174.0,P=0.000). The presence and aggravation of bone marrow edema indicate the enlargement of necrosis area and the aggravation of symptoms in patients with nontraumatic femoral head necrosis, which can be used as an auxiliary index for the appraisal of disease progression.

Objective To investigate the diagnosis and therapy of the sclerosing hemangioma of the lung.Mothods The case history of 15 patients were analyzed retrospectively and summarized the diagnosis and therapy of the Sclerosing hemangioma of the lung.Results There were 2 males and 13 females(1:6.5).No patients were diagnosed this disease before operation for there were no specific imageology features.The tumor cell indicated pantomorphia and there were 3 cases were misdiagnosed during the operation frozen section pathology.The optimization therapy methods was to resect the tumor with the VATS.The prognosis of this disease was well and there were no cases recurrence in these 15 patients.Conclusion The sclerosing hemangioma of the lung was the benign tumor and there were no specific imageology features.The misdiagnosis was frequently before operation.To resect the tumor with the VATS was the optimal therapy method.

Clinical data of 4 patients with chronic obstructive pulmonary disease who underwent allogenic single-lung transplantation were retrospectively analyzed. All cases received corpse donor lungs. One case with diffuse emphysema underwent right lung transplantation. and 3 received left lung transplantation. including one underwent fight lung volume reduction during surgery and 2 cases underwent right lung volume reduction post-transplantation. The inductive treatment with daclizumab or antithymocyte globulin was done and the rejection was prevented with Tacrolimus. mycophenolate and prednisone. The surgery was successfulin 4 cases. One case developed acute rejection on the fifth day post-transplantation. and controlled using methylprednisolone. Two cases discharged successfully, of whom one lived more than 2 years. Two cases died 74 days and 77 days after lung transplantation. respectively.

Objective To investigate the therapeutic effect of single stage cavernostomy and myoplasty on the aspergillus thoracic empyema. Method Four patients suffering from aspergillus thoracic empyema were treated with single stage cavernostomy and myoplasty. They got the inflammation of aspergillus after the operation for tuberculosis in 2, bronchiectasis in 1 and lung cyst in 1. They were performed rib resection, cavernostomy, closure of the bronchial openings and total/partial obliteration of the cavity with a muscle flap as a single stage technique. Pectoralis major muscle was used for the myoplasty in 1, latissimus dorsi in 1 and intercostal muscle in 2. All patients were given the therapy of nutritional support, local pressure dressing and oral anti-fungus drugs. Result Two patients got the primary healing, the chest tube were pulled out 2 days later and they were discharged 12 days after the operation. One patient′s cavity became little and there was pus drained from the chest tube, he cured after prolonged draining and washed with the anti-fungus drugs. One patient dead for pneumonia 67 days after the operation. Conclusion Cavernostomy and myoplasty as a single stage technique is safe and reliable in the management of patients with aspergillus thoracic empyema.

Ventilator aid respiration is important for patients with respiratory failure.But the negative emotional reaction of patient during connection and disconnection to ventilator will seriously influence the therapeutic effect,so a comprehensive underatanding about the emotional characteristic and dynamic emotion change of patient is of primary importance,because it will improve the diagnostic and therapeutic level,give patient personalized take care and make effective therapeutic plan,which will help keep a good emotion state for patient and accelerate recovery.This article will discuss how to use ventilator more effectively and economically.

Objective To report the experience of lung transplantation with size reduced graft lung.Methods Four cases receiving lung transplantation with size reduced graft lung were analyzed retrospectively.In case 1,left lung transplantation combined with contra-hteral lung volume mduction.In case 2,right lung transplantation Wag individually performed with partially msecfion of upper lobe of graft lung.In case 3.bilateral sequential lung transplantation wag performed using graft lung with partially resection of bilateral upper lobes.In the remained ease,bilateral sequential lung tansplantation was performed using graft lung with resection of right lower lobe.Results All the size reduced graft lungs had good functions during the peri-operation period.Case 1 and case 2 still survived without obvious complication.Case 3 experienced temporary air leak on the 5th day postoperation and cured by water seal drainage but died of abrupt bronchorrhea due to aspergillus infection on the 32th day postoperation.The last cage experienced smoothly recovery excepted fatal virus pneumonia 2 months postopemtion.Conclusion Size reduced graft lungs can be successfully used for transplantation.

OBJECTIVE To study the gene chip joint pyrosequencing technology in the newborn genetic deafness gene mutation screening, and provide a theoretical basis for the early diagnosis and prevention of genetic deafness. METHODS 2000 Neonatal EDTA umbilical cord blood was collected and genomic DNA (gDNA) was extracted. Microarray chip was used to detect four deafness gene at 9 mutation sites. And the positive result of gene chip detection was verified by pyrosequencing.RESULTS Among the GJB2 mutations, there were 1 case of 35delG mutation type, 3 cases of 176 del16 mutation type, 57 cases of 235del C mutation type, 9 cases of 299 del AT mutation type, 6 cases of GJB3 gene 538C>T mutation type. There were 5 cases of 1555A>G mutations and 1 case of 1494C>T mutations in mitochondrial 12S rRNA. There were 6 cases of 2168A>G mutation type and 23 cases of IVS7-2A>G mutations in SLC26A4. 103 cases of newborns carry the mutated gene in 2,000, the gene mutation rate is 5.15%. CONCLUSION All the four genes mutation at nine mutation sites are found in newborns with family history of non-hereditary deafness, and GJB2 gene mutation is common. The screening of genetic deafness in newborns is very important for early diagnosis and prevention of hereditary hearing loss. In particular, the diagnosis of mitochondrial 12S rRNA gene mutation can prevent the occurrence of deafness caused by drug use, for the genetic mutation of these carriers' health is of great significance.