OBJECTIVETo establish the method for cotransferring human A20 gene and human heme oxygenase-1 (HO-1) gene into the isolated rat islets using lentiviral transfection system, and to study the protective effect of A20 and HO-1 protein against the apoptosis induced by cycloheximide (CHX) and TNF-α, and finally to explore the underlying mechanism.

METHODThe A20 gene and HO-1 gene were cloned and inserted into the lentiviral transfection system. The efficacy of gene transfer was measured by the intensity of the enhanced green fluorescent protein (EGFP) fluorescence-positive islets. Western blot was applied to verify the expression of the A20 and HO-1 genes. To induce apoptosis in vitro, the isolated islets were treated with CHX+TNF-α, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and the fluorescence-activated cell sorting (FACS) methods were used to evaluate the apoptosis of the islet cells and Western blot was used to detect caspase-3 activation.

RESULT(1) A20 and HO-1 genes were introduced into the isolated islets by lentiviral transfection, both of the genes were highly expressed in the islets after 96 hours culture detected by Western blot method. (2) The insulin levels in the cell culture medium from A20 and/or HO-1 transgenic islets were significantly higher than that in non-transgenic controls (P < 0.01). (3)After CHX + TNF-alpha treatment, the cell culture medium insulin concentration in the A20 gene transfected group [(93.58 ± 4.12)µg/ml], HO-1 gene transfected group [(88.98 ± 4.77) µg/ml ] and A20/HO-1 co-transfected group [(103.33 ± 3.16) µg/ml] were significantly higher than that in the EGFP group [(9.03 ± 0.65) µg/ml ] and the control group [(8.86 ± 0.38) µg/ml] (P < 0.001). Minimum expression level of the activated caspase-3 was found in the A20/HO-1 co-transfected group.

CONCLUSIONThe lentiviral gene transfer system was an efficient and stable gene transfer vector, the over-expressed A20 and HO-1 protein delivered via lentivirus could preserve rats' islets function and act against the apoptosis induced by CHX and TNF-α.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Flow Cytometry ; Genetic Vectors ; Heme Oxygenase-1 ; genetics ; metabolism ; Humans ; Insulin ; metabolism ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Islets of Langerhans ; drug effects ; enzymology ; physiology ; Lentivirus ; genetics ; Male ; Nuclear Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection ; methods ; Tumor Necrosis Factor alpha-Induced Protein 3 ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo evaluate clinical results of an interspinous stabilization system (Wallis) in treating lumbar degenerative disease in the short-term.

METHODSFrom August 2007 to June 2010,48 patients with lumbar degenerative disease who were treated with interspinous stabilization system, the data of patients were analyzed retrospectively. In all of the 48 cases, there were 30 males and 18 females with an average age of 54.2 years (ranged, 40 to 68 years). Forty-four cases were with single segment and 4 cases with two segments. Of them, 4 cases were in L3, 4, 40 cases were in L4, 5, 4 cases were in L3, 4 and L4, 5. The radiographic data of patients were analyzed. Clinical effects were evaluated by Japanese Orthopedic Association (JOA) score system and low back pain disability questionnaire (Oswestry) and Odom method.

RESULTSAll the patients were followed up from 1 to 2 years with an average of 18 months. According to Odom's criteria, 20 cases obtained excellent results, 24 good, 4 fair. JOA score increased from 12.4 +/- 2.7 preoperatively to 26.1 +/- 2.0 postoperatively (P < 0.01). Oswestry score decreased from 14.1 +/- 2.9 preoperatively to 5.5 +/- 1.8 postoperatively (P < 0.01). The posterior height of intervertebral space and height of nerve root canal increased compared with that of preperative height.

CONCLUSIONThe treatment of lumbar degenerative disease with interspinous stabilization system can obtain satisfactory effects in the near future. It can retain dynamic stable of corresponding segments, expand volume of vertebral canal, and is safe and feasible.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Stenosis ; surgery

OBJECTIVETo develop a simple, rapid and reliable method of purifying Sprague-Dawley (SD) rat islets by sequential filtration through two cell strainers of different sizes and to evaluate the efficacy of the method.

METHODSIslets were isolated from 8 to 12-week-old clean grade Sprague-Dawley rat pancreases using the standard collagenase digestion procedure and purified with either the generally used Ficoll density gradient method or the innovative two-step sequential filtration method. The purity and vitality of the isolated islets were visualized and assessed with DTZ and AO/PI staining. Glucose stimulating tests were performed to assay cell activity, and immunohistochemical staining was used to evaluate the synthesis function of islet cells.

RESULTSThe yield of islets in the two-step filtration method group was 782±115 IEQ per rat, which was significantly higher than in the conventional Ficoll density gradient method group (598 ± 135 IEQ per rat, P < 0.01). Purity of the isolated islets in the two-step filtration method group was 90%-100% and vitality was over 95%. In the conventional Ficoll density gradient method group, islet purity was 65%-85% and vitality was 85%-95%. With regard to the high-sugar stimulation test in the two-step filtration method group, insulin concentrations in islets cultured for 24 hours were significantly higher than in those that were freshly purified (76.9 ± 6.1 μg/L vs 49.4 ± 3.9 μg/L; P < 0.01).

CONCLUSIONSA two-step sequential filtration method for rat islet purification was developed and the method was simple and reliable, with high islet vitality, purity and yield.

Animals ; Cell Separation ; methods ; Female ; Filtration ; Immunohistochemistry ; Insulin ; biosynthesis ; Islets of Langerhans ; cytology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo analyze the intermediate-term results associated with the use of Zweymtiller hip system.

METHODSReview the 116 cases (142 hips) who were treated with total hip replacement from 1996 to 2002 by a single surgeon using cementless Zweymüller hip systems.

RESULTSSixty-one cases (77 hips) were followed up, 50 cases (64 hips) were evaluated both clinically and radiographically while 5 cases (6 hips) and 6 cases (7 hips) were only evaluated clinically and radiographically respectively. The average follow-up period was 7.3 years (range 5 to 11 years). The mean preoperative Harris Hip Score was 46 while the mean postoperative Harris Hip Score was 93. Distal cortical hypertrophy and medullary sclerosis were observed in 30 hips (42.3%) and 33 hips (46.5%) respectively. Heterotopic ossification arose in 45 hips (63.4%). Radiolucent lines occurred in 27 stems (38.0%) but in no cups. Radiolucent lines were mostly observed in Gruen zones 1. Osteolysis occurred in 7 cups (9.9%) and 18 stems (25.4%). Osteolysis was mostly observed in Delee Zone 3 and Gruen zone 7. In the distal Gruen zones 3, 4 and 5, no radiolucent line or osteolysis was observed. No hips had been revised, 3 cups needed revision surgery because of aseptic loosening.

CONCLUSIONThe 5-11 years follow-up results are satisfactory, but osteolysis is common.

Adolescent ; Adult ; Aged ; Arthroplasty, Replacement, Hip ; Female ; Follow-Up Studies ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

italic>Schisandra chinensis is a traditional Chinese medicine with the functions of reinforcing deficiency, strengthening, and inducing astringency, appliable to treat the chronic cough and deficiency in breath, palpitation, and insomnia, etc. A hybrid mass spectrometry scanning strategy (high-definition data-independent/data-dependent acquisition, HDDIDDA), enabling the ion mobility separation and alternating data-independent acquisition/data-dependent acquisition, was established, which, in combination with in-house library-driven automatic peak annotation workflows facilitated by the UNIFI software, was utilized to systematically characterize the multi-classes of chemical components from S. chinensis. The use of an HSS T3 column (100 mm × 2.1 mm, 1.8 μm), 0.1% formic acid in H₂O-acetonitrile as the mobile phase running at the flow rate of 0.3 mL·min^-1, and column temperature at 35 ℃, could enable good separation of the S. chinensis components within 42 min. HDDIDDA scan in both the positive and negative ion modes was employed for data acquisition. Based on the automatic peak annotation, reference standards comparison, MS² data interpretation, and literature analysis, we were able to identify or tentatively characterize 105 compounds in the S. chinensis decoction, involving 56 terpenoids, 42 lignans, five glycosides, one organic acid, and one flavonoid. HDDIDDA scanning can improve the coverage of data acquisition and improve the accuracy of identification, while CCS prediction analysis provides the possibility to distinguish isomers by the ion mobility technology. The results provide reference for the intelligent material basis research of TCM.

Objective To explore the relationship between the radiological characteristics and clinical and pathological manifestations of intracranial juvenile xanthogranuloma. Methods The radiological and clinical characteristics of two intracranial juvenile xanthogranuloma cases were analyzed respectively. In combination with reviewing literatures, the radiological characteristics and discipline of intracranial juvenile xanthogranuloma disease were also analyzed. Results Intracranial juvenile xanthogranuloma disease was accidently detected in one case. The other case was detected with headache and numbness in face. No abnormal sign was found in skin. On plain CT isodensity signals appeared and on contrast-enhanced CT homogeneous enhancement was found.On MR imaging,masses showed isointense signals on T1WI and iso-or hypo-intensity signals on T2WI, and enhanced homogeneously. Lesions were solitary or multiple in the external of brain. After operation intracranial juvenile xanthogranuloma was confirmed by pathology, and no recurrence occurred. Conclusion Intracranial juvenile xanthogranuloma is a kind of rarely encountered disease with typically radiological characteristics, which is difficult to detect before surgery and can be confirmed by post-operative pathological and immunohistochemical examinations.[Chinese Medical Equipment Journal,2018,39(5):71-74]

OBJECTIVETo evaluate the early clinical results of two stage hip replacement after failed internal fixation for femoral neck fractures in young patients.

METHODSFrom June 2008 to June 2010,24 patients with femoral head necrosis caused by failed internal fixation were treated with hip arthroplasty. Among them, 12 patients were male and 12 patients were female, with an average age of 42.9 years old (ranged, 18 to 58). According to Harris score and X-ray examination, the clinical result was evaluated.

RESULTSTwenty-three cases were followed up with an average age of 34.4 months (ranged, 25 to 48). After operation, 1 case complicated with the dislocation of hip joint. No deep infection of hip joint, prosthetic loosing or peripheral fracture was found. The mean Harris score was 90.9 +/- 4.3, and 18 obtained excellent results, 4 good and 1 fair.

CONCLUSIONAlthough treatment of femoral head necrosis with two stage hip replacement after failed internal fixation is difficult during operation, its early result is satisfactory.

Adolescent ; Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femoral Neck Fractures ; surgery ; Femur Head Necrosis ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Failure

Objective To investigate the clinical presentation,diagnosis,treatment and outcome of patients with pulmonary veno-occlusive disease (PVOD).Methods Data from patients diagnosed as PVOD from May 2008 to May 2011 in Shanghai Pulmonary Hospital,Tongji University were retrospectively reviewed.Results During this period,5 patients [ 4 female,aged from 12 to 42 (22 + 12 ) years old ] were diagnosed as PVOD.The durations from symptoms onset to PVOD diagnosis was 2 to 50 ( 16 ± 20)months and four of them were previously diagnosed as idiopathic pulmonary arterial hypertension.All patients at the time of PVOD diagnosis had a severely impaired WHO pulmonary hypertension functional class (3 in class Ⅲ and 2 in class Ⅳ ).Furthermore,all patients characterized by a typical sign of centrilobular ground-glass opacities in high-resolution computed tomography,a markedly reduction of diffusing capacity of the lung for carbon monoxide [ (38 ± 12)％ of predicted value] in pulmonary functional test and severely compromised cardio-pulmonary hemodynamics identified by right heart catheterization.All patients received conventional and pulmonary arterial hypertension specific therapies,and then followed-up regularly.Up to now,4 out of 5 patients died due to refractory right heart failure.The durations from symptoms onset to death and from PVOD establish to death were 5 - 65 ( 27 + 26 ) months and 1 - 16 (9 ± 9 ) months,respectively.Conclusions PVOD is a rare and malignant cardio-pulmonary disorder that often be misdiagnosed as idiopathic pulmonary arterial hypertension.Given the poor responses to modem pulmonary arterial hypertension specific therapies,lung transplantation remains the treatment of choice.

OBJECTIVETo analyze the clinical, imaging and pathological findings of congenital intrapulmonary lymphangioma and hemangioma in 5 infants and young children.

METHODData of 3 cases with congenital intrapulmonary lymphangioma and 2 cases with haemangioma were analyzed.

RESULTAll the 5 cases had cough, difficulty in breathing, cyanosis of lips, and shortness of breath. Imaging study indicated cystic and space occupying changes of partial lung. In the two cases of hemangioma, the blood vessels passing through the hemangioma were observed on CT films. Histochemical studies showed that the cystic pockets from removed tissues were different in size and there was a line of flat endothelial tissue around these pockets. Immunochemical studies indicated D2-40 positive, factor VIII weak positive for lymphangioma cases; while in hemangioma cases, factor VIII was positive, D2-40 was negative or weakly positive, and both of cytokeratin was negative.

CONCLUSIONIt is very important to consider and identify congenital intrapulmonary lymphangioma or hemangioma when a patient has cystic pockets and space occupying change in their lung by imaging and pathological studies.

Female ; Hemangioma ; congenital ; pathology ; Humans ; Infant ; Lung Neoplasms ; congenital ; pathology ; Lymphangioma ; congenital ; pathology ; Male

OBJECTIVETo investigate the incidence, predisposing factors and therapeutic modalities of acute colonic pseudo-obstruction (ACPO) in patients after total hip arthroplasties (THA) and total knee arthroplasties (TKA).

METHODSFrom January 2006 to December 2009, 12 patients with ACPO after THA and TKA operation were investigated retrospectively,who were viewed as the ACPO group. There were 10 males and 2 females with an average age of (78 +/- 12) years in the ACPO group. Other 853 patients without ACPO after THA and TKA operation were viewed as the control group treated at the same period. The incidence of ACPO was calculated. The clinical data were collected and compared between the two groups including patient age, gender, procedure, anesthetic class, clinical presentation, radiographic findings, duration from index surgery to diagnosis of ACPO, treatment, postoperative mobilization time, and length of hospital stay.

RESULTSThe incidence of ACPO was 1.4%. The incidence of primary THA (1.3%) was higher than that of primary TKA (0.4%); the incidence of hip and knee revisions (5.0%) was higher than that of primary THA and TKA (1.0%); there was no difference in incidence between hip revisions (5.5%) and knee revisions (4.0%). The mean age was (78 +/- 12) years old in ACPO group and (71 +/- 13) in the control group. The male/female ratio was 5:1 in ACPO group and 2:3 in control group. There were statistical differences in mean age and gender ratio between the two groups. No association was found with respect to anesthetic class. On average, ACPO occurred at 2.5 days after index surgery. The abdominal distention occurred in all 12 cases, nausea or vomiting in 8 cases and abdominal pain in 3 cases. Radiographically cecal dilation occurred in all cases and intestinal dilation in 3 cases. All patients initially were treated conservatively with immediate cessation of oral intake,a nasogastric tube and oral mineral oil. Three patients received a rectal tube. Only 1 patient required endoscopic decompression. There were no deaths after ACPO in the series. Mean mobilization time after surgery averaged (5.0 +/- 2.2) days in ACPO group compared with (2.5 +/- 1.1) days in the control group. Mean hospital stay averaged (16.5 +/- 6.4) days in ACPO group compared with (10.5 +/- 4.5) days in the control group. There were statistical differences in mean mobilization time after surgery and mean hospital stay between two groups.

CONCLUSIONACPO mainly happened in old male patients. The majority cases response to conservative treatment and their prognoses are good. But ACPO will delay mobilization time after surgery and increase hospital stay.

Acute Disease ; Aged ; Arthroplasty, Replacement, Hip ; adverse effects ; Arthroplasty, Replacement, Knee ; adverse effects ; Case-Control Studies ; Colonic Pseudo-Obstruction ; diagnosis ; epidemiology ; etiology ; therapy ; Female ; Humans ; Incidence ; Male ; Middle Aged