Objective · To investigate the expression of the regulator of calcineurin 1 (RCAN1) and calcineurin A (CnA) in tissues of in-stent restenosis after intervention of arteriosclerosis obliterans (ASO), and to explore the relationship between their expression levels and the occurance of in-stent restenosis. Methods · Superficial femoral arterial tissues were collected from 15 ASO patients undergoing lower extremity amputation for in-stent restenosis in Department of Vascular Surgery, The First Affiliated Hospital of Chongqing Medical University from September 2013 to June 2016. H-E staining and Masson staining were performed on the stenosis tissues, as well as on the proximal and distal tissues, and the morphological changes of these tissues were observed under optical microscope. Western blotting was used to detect the protein levels of RCAN1, CnA and proliferating cell nuclear antigen (PCNA). The distribution of RCAN1 and CnA proteins was observed by immunohistochemistry and immunofluorescence methods. In addition, co-immunoprecipitation was used to validate the protein-protein interaction between RCAN1 and CnA in vascular tissues. Results · The expression of RCAN1 in the distal tissues was significantly elevated compared with the proximal tissues and the stenosis tissues (P<0.05). The expression of RCAN1 in the proximal tissues was higher than that in the stenosis tissues (P <0.05). The expression of CnA and PCNA in the stenosis tissues was significantly elevated compared with the proximal tissues and the distal tissues (P<0.05). Immunohistochemistry and immunofluorescence analyses showed that RCAN1 and CN proteins were mainly expressed in the cytoplasm of vascular smooth muscle cells. Co-immunoprecipitation analysis showed there is protein-protein interaction between RCAN1 and CnA in arterial tissues. Conclusion · The low expression of RCAN1 and the high expression of CnA are probably related to the occurrence of in-stent restenosis.

Objective To investigate the long-term efficacy of nonmyeloablative autologous peripheral blood stem cell transplantation(NAST) to cure refractory autoimmune disease(AD).MethodLong-term follow up of four cases of AD patients with NAST were summarized.The pretreatment regimen was intravenous injection of cytarabin (200 mg· kg-1· d-1 ) and cyclophosphamide (40 mg· kg-1· d-1).The therapeutic effect was evaluated by the change of symptoms and signs and long term complications.Changes of immune function were detected by flow-cytometry.ResultsFive cases of patients had been successfully engrafted.The average time for peripheral leucocytes count to reach 4.0×109/L was 12 days.It needed 10 days for platelets to return to 100×109/L and 22 days for hemoglobin to 120 g/L.Apparent remission of symptoms and signs was observed after transplantation.Lymphocyte subtypes analysis pre- and post- NAST showed that count of CD4+ and the ratio of CD4 +/CD8 + was returned to normal.One patient gave birth to a healthy baby four years after transplantation.Three female patients returned tonormal life. Conclusions Compared with classical myeloablative stem cell transplantation,NAST has a rapid hematopoietic recovery and good long-term therapeutic effect in AD.The quality of life in AD patients treated with NAST is higher than those treated with myeloablative hematopoietic stem cell transplantation.

Objective To compare the clinical efficacy and safety of two different conditioning regimens in nonmyeloablative autologous peripheral blood stem cell transplantation (NAST) for the treatment of systemic lupus erythematosus (SLE).Methods Different conditioning regimens were used in two groups:cytarabin combined cyclophosphamide in group 1 and ATG combined cyclophosphamide in group 2.Different recovery time of leucocytes,neutrophils and platelets in the two groups were compared.Statistical analysis were carried out by paired t-test.Results The mean time for peripheral leucocytes reaching 1.0×109/L,neutrophils getting up to 0.5×109/L,platelet raising to 100×l09/L and hemoglobin rising to 120 g/L in group 1 were [(7.2±1.3),(8.0±1.5),(10.5±1.4),(22.1±2.3)days] and [(10.4±2.1),(12.0±1.9),(19.3±2.1),(28.1± 2.4)] days in group 2.The difference was statistically significant (P＜0.01).CD4+ cell count and the ratio of CD4+/CD8+ of pre- and pro-NAST was changed.No significant differences were observed in the two groups.Conclusion For the sake of safety and hematopoietic reconstitution,we recommend cytarabin combined cyclophosphamide as the preferred conditioning regimen.

Staphylococcus aureus ( S. aureus ) osteomyelitis, a significant complication for patients un-dergoing fracture fixation, is a great challenge for orthopaedic surgeons due to its extreme difficulty in mangae-ment. Animal models play an important role in exploring the pathogenesis of osteomyelitis and determining the efficacy of prophylactics and therapeutic treatment. To help understand current animal models of S. aureus os-teomyelitis, we conduct a systematic search to identify animal experiments that have investigated the management of S. aureus osteomyelitis. Experimental studies are categorized by animal species and are further classified by the setting of infection. Study methods are summarized and the advantages and disadvantages of each species and model are discussed.

Objective To study the anti-fatigue effect of Cynomorium songaricum flavone(CSF)on old rats.Methods50 old male Wistar rats were randomly divided into the control group(no swimming,no drugs),single swimming group(swimming,given saline by gavage),and three swimming plus CSF groups(swimming,given CSF 20 g/kg weight,10 g/kg weight and 5 g/kg weight respectively),total five groups with 10 animals in each group.Animals swam with free swimming in endless cycles water 30 minutes once every day for 10 days.The level of monoamine oxidase(MAO) and glutathione peroxidase(GSH-px) in blood of animals were examined by MAO kit,GSH-px kit and ultraviolet spectrophotometer at 11th day.ResultsCompared with single swimming group,the body weights of swimming plus CSF groups increased significantly by 12.9%,80.6% and 200% respectively(P<0.01);swimming times before sinking increased by 19.4%,29% and 48.4%;sinking times of each rat(time/10 min) decreased by 17.8%,43.2% and 72.9%;total swimming times increased by 2.8%,7% and 29.6% respectively(P<0.01);MAO decreased by 60%,69.8% and 80.5%,GSH-px increased by 465.3%,563.8% and 635% respectively(P<0.01).ConclusionCSF shows anti-oxidative and anti-fatigue properties and can be given as prophylactic/therapeutic supplements for increasing antioxidant enzyme activities and preventing lipid peroxidation during strenuous exercise.

Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis (FSGS) and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as healthy control group. Genomie DNA was extracted from peripheral blood cells of FSGS patients and hair of patients' parents by polymerase chain reaction (PCR) and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database (www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation, corn). Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basie vector, pRL-SV40 and PCI2 cell line. Hair DNA of novel mutation patients' parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients' kidney tissue was examined by immunofluorescence. Results Three patients with 1-34C>T, 1-590delA and (1-1044delT)+ (I-797T >C) +(1-769A >G) heterozygous mutations were found in ACTN4 gene promoter respectively, and two patients with 1-24G>A and 1-851C>T heterozygous mutations in SYNPO gene promoter respectively. The same mutations were not found in the control group of 70 healthy people. Except one patient accepting her parents' 1-1044delT and 1-797T>C mutated chromosome respectively, no same mutations were found in patients' parents. Protein expression of alpha-actinin-4 and synaptopodin was reduced in mutated patients' kidneys. Except 1-1044delT group, luciferase activity in mutated groups decreased. (1-1044delT)+(1-797T>C)+(1-769A>G) mutation was associated with poor outcome and patient with these mutations progressed to end-stage renal failure. Conclusion Mutations of ACTN4 and SYNPO gene promoters affect gene transcription and protein translation, which may contribute to the onset of sporadic primary FSGS.

Objective To summarize the outcome of nasopharyngeal carcinoma ( NPC) treated by helical tomotherapy in the Chinese PLA general hospital. Methods Between September 2007 and August 2010, 121 newly diagnosed NPC patients were treated by radiotherapy with Tomotherapy system, with ( n =90) or without ( n = 31) concurrent chemotherapy or molecular target therapy. The prescription dose was 70 - 74 Gy/33f to primary tumor and positive lymph node planning target volume,60. 0 - 62. 7 Gy/33f to high risk planning target volume, and 52 -56 Gy/33f to low risk planning target volume. Acute side-effects were evaluated with RTOG/EORTC criteria. Results The remission rate of primary lesion and positive lymph nodes was 95. 0％ and 99. 0％ , respectively. The follow-up rate was 100％ . The number of patients with 1 ,2 and 3 years followed-up were 99 , 49 , and 7. The 1-, 2-and 3-year local relapse-free survival rates were 97. 30％ , 97. 3％ and 97. 3％ , respectively. The 1-,2-and 3-year nodal relapse-free survival rates were 100％ , 100％ and lOO％, respectively. The 1-, 2-and 3-year distant metastasis-free survival rates were 98. 4％ , 96. 3％ and 96. 3％ , respectively. The 1-, 2-and 3-year overall survival rates were 96. 5％ ,92. 6％ and 86. 8％ , respectively. Acute toxicities of skin, oral mucosa and xerostomia with grade 0,1,2 and 3 were 5. 0％ , 74. 4％ , 15. 7％ and 4. 9％ ; 0. 8％ , 37. 2％ , 57. 9％ and 4. 1％ ; 3. 3％ , 53. 7％ ,43. 0％ and 0％ , respectively. Xerostomia restored with time, no grade 2 or more xerostomia was observed 1 year after radiation therapy. Concurrent chemotherapy significantly increased incidence of mucositis,esophagitis and tracheitis. Conclusion Helical tomotherapy is efficient, secure and effective modality for the treatment of nasopharyngeal carcinoma.

The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism

Objective To evaluate the role of Toll-like receptor 4 (TLR4) signaling pathway in sevoflurane anesthesia-induced cognitive dysfunction in aged rats.Methods Twenty-seven SPF healthy male Sprague-Dawley rats,aged 20 months,weighing 550-750 g,were divided into 3 groups (n =9 each) using a random number table:control group (C group),sevoflurane anesthesia group (S group) and TLR4 antagonist plus sevoflurane anesthesia group (TS group).TLR4 monoclonal antibody 30 μl was injected into the lateral cerebral ventricle in group TS,and the equal volume of serum containing no antibody was injected into the lateral cerebral ventricle in C and S groups.At 10 min after completion of injection,S and TS groups inhaled the mixture of 4％ sevoflurane and 30％ oxygen for 6 h,and group C only inhaled the mixture of air and oxygen.Morris water maze test was performed at 24 h after the end of sevoflurane anesthesia.The animals were sacrificed after completion of Morris water maze test,brains were removed and hippocampi were isolated for determination of nerve cell apoptosis (using TUNEL) and expression of activated caspase-3 (using immunofluorescent staining).Nerve cell apoptosis rate was calculated.The expression of high-mobility group box 1 protein (HMGB1) mRNA in hippocampi was measured by Northern blot assay at 6 h after the end of sevoflurane anesthesia.The expression of amyloid precursor protein (APP) and amyloid beta protein (Aβ) in hippocampi was assessed by Western blot at 24 h after the end of sevoflurane anesthesia.Results Compared with C group,the escape latency was significantly prolonged,nerve cell apoptosis rate was increased,the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was up-regulated in group S,and nerve cell apoptosis rate was increased,the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was up-regulated (P＜0.05),and no significant change was found in the escape latency in group TS (P＞0.05).Compared with S group,the escape latency was significantly shortened,nerve cell apoptosis rate was decreased,and the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was down-regulated in group TS (P＜0.05).Conclusion Activation of TLR4 signaling pathway is involved in the mechanism of sevoflurane anesthesia-induced cognitive dysfunction in aged rats.

Objective To compare the efficacy of CT-guided radiofrequency thermocoagulation (RFTC) treatment through the inside and outside of the foramen rotundum for primary trigeminal maxillary neuralgia.Methods Forty-eight patients of both sexes,aged 58-75 yr,suffering from primary trigeminal neuralgia the 2nd trigeminal branch pain,scheduled for elective CT-guided trigeminal RFTC,were divided into 2 groups (n =24 each) using a random number table:inside of foramen rotundum group and outside of foramen rotundum group.The needle was inserted until the lateral 1/3 of the foramen rotundum was reached under CT guidance in inside of foramen rotundum group or until the outside of the foramen rotundum was reached in outside of foramen rotundum group.High-temperature RFTC 95 ℃ was performed for 120 s,repeating for 2-3 cycles.The current sensory threshold at different frequencies was measured at 1 day before and after operation.Numeric rating scale score was performed at 1 day,30 days,1 yr and 2 yr after operation,effective RFTC treatment was defined as numeric rating scale score≤ 1,and the condition of postoperative effective RFTC treatment was recorded.The development of adverse reactions such as facial hematoma,corneal ulcer,injuring the other branch and cerebrovascular accidents was recorded.Results Compared with outside of foramen rotundum group,the current sensory threshold on the affected side at frequency of 2 000 Hz was significantly increased at 1 day after operation,the rate of effective RFTC treatment was increased 2 yr after operation (P＜0.05),and no significant change was found in the incidence of postoperative facial hematoma in inside of foramen rotundum group (P＞0.05).Conclusion The long-term curative effect of CT-guided RFTC treatment for primary trigeminal maxillary neuralgia through the inside of the foramen rotundum is better than that through the outside of foramen rotundum.