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Objective@#To investigate GATA3 expression and the regulatory mechanism of m6A modification in the re- sponse of alveolar epithelial cells to radiation, and to provide a new therapeutic target for radiation-induced lung injury based on its pathogenesis.@*Methods@#Human lung epithelial cell line (A549) and mouse lung epithelial cell line (MLE-12) were exposed to X-ray irradiation with a single dose of 10 Gy (dose rate 1 Gy/min) and 6 Gy (dose rate 0.75 Gy/min), respect- ively. The expression of VIRMA gene (RNA methylase) was inhibited by lipofection of A549 cells and MLE-12 cells with shRNA-VIRMA plasmid and siRNA-VIRMA interfering fragment, respectively. Quantification of m6A RNA methylation was performed by colorimetry. Changes in the expression of mRNAs of VIRMA, GATA3, and epithelial-mesenchymal transition (EMT) markers in irradiated A549 and MLE-12 cells were determined by qRT-PCR. Changes in the expression of VIRMA, GATA3, and EMT marker proteins in irradiated A549 and MLE-12 cells were determined by Western blot.@*Results@#Radiation up-regulated the expression of methylase VIRMA in A549 and MLE-12 cells, which in turn enhanced the m6A of total RNA and the expression of GATA3 gene and protein, resulting in EMT. Furthermore, in A549 and MLE-12 cells, interference of the VIRMA gene significantly reduced the expression of GATA3 gene and protein and the expression of EMT-related molecules.@*Conclusion @#Radiation induces m6A modification in alveolar epithelial cells, which up-regu- lates the expression of GATA3 gene and induces EMT, thus playing an important role in the process of radiation-induced lung injury.
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Objective:To explore the surgical technique and clinical effect of pressure boost in repairing soft tissue defects of limbs with thinned anterolateral thigh perforator flap (ALTP) .Methods:From January, 2015 to December, 2018, 18 cases with soft tissue defects of limbs with various damages of blood vessels and nerves with explosure of tendon and bone. There were 13 males and 5 females aged between 18 to 56 (averaged of 36.3) years, which were 6 defects in shank, 4 in foot and ankle, 5 in forearm, and 3 in hand. The soft tissue defect area was 7 cm ×12 cm to 13 cm ×30 cm. Thinned ALTP was used to repair the wound surface. The perforating vessels of the distal flap were anastomosed with one branch of the internal vessel pedicle flap to increase the pressure hence the blood supply of the distal region. The donor sites were sutured directly or covered by skin graft. Followed-up was conducted by 1-2 monthly clinic visits and telephone or on-line review to check the flap survival and recovery of functions.Results:All flaps survived without arterial or venous crisis. One flap had partial necrosis at the distal end, and healed after dressing change. One case had a swelling flap due to a congestion beneath the flap. The wound achieved primary healing after removal of sutures, ligation of subcutaneous vessels and drainage of hematoma. All patients were followed-up for 6 to 18 (average, 9.5) months. All flaps had good appearance and texture. After rehabilitation treatment, most of the joint activity had been recovered: extension and flexion of wrists joints ranged 60°-80°, 70°-80° for metacarpophalangeal joints and 40°-60° for ankle joints. One patient underwent ankle joint dorsiflexion function reconstruction and flap thinning at 6 months after operation due to the defects of most of the extensor tendon.Conclusion:During the use of free ALTP to repair soft tissue defect of limbs, application of the technique of pressure boost is able to increase blood supply to the distal region of flap. It helps to reduce the incidence of infection and necrosis at the edge of the flap.
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Objective To investigate and explore the clinical value of single-photon emission computed tomography(SPECT) imaging in the differential diagnosis of neonatal obstructive jaundice.Methods From January 2012 to April 2017,130 cases of suspected neonatal obstructive jaundice in the First Hospital of Jiaxing were enrolled in the study.All patients were given SPECT hepatobiliary scintigraphy,were injected 99mTc-EHIDA imaging agent,99mTc-MIBI imaging agent,diagnosis of hepatobiliary scintigraphy after the acquisition.The result of operative cholangiography was viewed as the gold standard,the diagnostic sensitivity,specificity,accuracy of simple 99mTc-EHIDA SPECT imaging examination,combined with SPECT imaging agent in the diagnosis of neonatal obstructive jaundice were calculated and compared.The coincidence rate of differential diagnosis of different etiologies of neonatal obstructive jaundice was also compared.Results The diagnostic sensitivity,specificity and accuracy of combined with SPECT imaging agent in the diagnosis of neonatal obstructive jaundice were 94.23％,96.15％,94.61％,respectively,which of simple 99mTc-EHIDA imaging agent SPECT were 83.65％,76.92％,82.31％,respectively,the differences were statistically significant(x2 =5.915,4.127,9.646,all P ＜ 0.05).The coincidence rates of extrahepatic biliary atresia,hepatitis syndrome of the two kinds of SPECT imaging agents combined examination were 96.05％,96.43％,respectively,which were significantly higher than those of the 99mTc-EHIDA imaging agent SPECT examination (84.21％,85.00％) (x2 =5.991,5.250,all P ＜ 0.05).Conclusion SPECT imaging in the diagnosis of neonatal obstructive jaundice has high sensitivity and accuracy,especially in combination with 99mTc-EHIDA imaging agent,99mTc-MIBI imaging agent can identify and distinguish different causes of jaundice,and provide a reliable basis for clinical treatment.
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The unique solution-gel transition property of in-situ gel makes it have advantages of good histocompatibility, long residence time, high local concentration, promising bioavailability and so on.This paper summarized the different types and the latest research progress in Chinese medicine targeting preparations of in-situ gel in order to provide reference for the application of in-situ gel in Chinese medicines.
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Objective To test whether multiplex ligation-dependent probe amplification(MLPA)could be used for the prenatal detection of the most common aneuploidies of chromosomes 13,18,21,X,and Y.Methods 34 cases including 22 blood samples(12 with trisomy 21,1 with monosomy X,one male witll extra Y and 8 healthy persons),4 cord blood samples with Down syndrome and 8 amniotic fluid samples ( 1 with trisomy 21 and 7 normal fetuses)were recruited into this study.All samples were confirmed by karvotype analysis. DNA was extracted from blood and amniotic lysate was incubated with proteinase K.MLPA was used to determine the relative copy numbers.Results The resuhs were available within 48 h and were concordant with karyotype analysis in all but one case of amniotic fluid that was suggested to be triploid sample 69,XXY by MLPA or contaminated by maternal blood.This sample actually was found containing a number of red blood cells after centfifugation in test. In total,the concordance rate with clinical characteristics was 97.1%.The Ratio values of 13,18,21,X in normal samples were approaching 1.0 except chromosome Y having slightly higher variation in relative copy number.The difference of ratio means between the normal and trisomy 21 samples was statistically significant by one-way ANOVA(F=298.906.P=0.000).Conclusion Computer assisted MLPA with high sensitivity is a rapid,simple,automatic and reliable method for detection of common chromosomal aneuploidies.
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BACKGROUND: Uncoupling of major histocompatibility complex (MHC) is the main cause for transplantation rejection, and it is the best way to prevent transplantation rejection by induce immunological tolerance of the recipient to the donor organ. Self-tolerant T cells can be obtained by negative selection in thymus, whether the intrathymic injection of allogenic antigen can get the immunological tolerance to the antigen?OBJECTIVE: To observe the effects of intrathymic injection of allogenic antigen on inducing immunological tolerance in nerve transplantation.DESIGN: A comparative observation.SETTINGS: Department of Immunology, Harbin Medical University; Department of Orthopaedics, Fourth Clinical Hospital of Harbin Medical University.MATERIALS: Thirty donor C57BL/6 mice (H-2b), male, aged from 6-8 weeks, weighing 18-22 g, were purchased from the Veterinarian Institute of Heilongjiang Province; While 60 recipient Balb/c mice (H-2b) female, aged from 6-8 weeks, weighing 18-22 g, from Beijing Experimental Animal Center. MHC (H-2b) antigen was prepared by the Department of Immunology, Harbin Medical University, and the concentration of protein was 4.4 g/L.METHODS: The experiments were carried out in the Department of Immunology, Harbin Medical University from June to November 2002. The recipient Balb/c mice were randomly divided into four groups: intrathymic injection group, syngenic transplantation group, allogenic transplantation group and immunosuppressant drug group. MHC (H-2b) antigen was extracted from splenic cells of donor C57BL/6 mice and injected intrathymically into recipient Balb/c mice (H-2d). Two weeks later, the sciatic nerve was transplanted to the recipient mice. Mixed lymphocyte reaction and delayed-type hypersensitivity (DTH) were detected at 3 weeks after transplantation.MAIN OUTCOME MEASURES:The differences of mixed lymphocyte reaction and DTH were compared among the groups.RESULTS: All the 30 donor C25BL/6 mice (H-2b) and 60 recipient Balb/c mice (H-2d) were involved in the analysis of results.①Results of mixed lymphocyte reaction: The cell proliferations in the syngenic transplantation group and intrathymic injection group were obviously lower than that in the allogenic transplantation group [(546.1±75.1), (2 668.3±533.8), (3 101.3±429.1), (4 312.3±534.1) minutes-1, P＜0.05].②Results of DTH: The thicknesses difference between two pads in the syngenic transplantation group and intrathymic injection group were obviously lower than that in the allogenic transplantation group [(41.1±3.7), (72.1±5.1), (57.6±11.3), (86.2±13.2)μm, P＜0.05].CONCLUSION:The intrathymic injection of donor H-2b antigen could induce immunological tolerance of nerve transplantation.
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Objective:To investigate the effect of intrathymic injection of allogenic antigen to sciatic nerve transplantation.Methods:C57BL/6(H-2b) mice were used as donors and BALB/c(H-2d) as recipients. The recipients were divided into four groups: auto-transplantation group, allogenic transplantation group, allogenic transplantation and using immunosuppressive drugs,intrathymic injection group: 3 mol/L KCl MHC antigen extractions were injected into the recipients’ thymus before two weeks before the sciatic nerves were transplanted. In the third week all the recipients underwent immunological detections for IL-2R,TNF-?,mixed lymphocytes culture and apoptosis.Results:All the detections indicated that it was of significant difference between intrathymic injection group and those in allogenic transplantation group.Conclusion:The immunological rejection of allogenic peripheral nerve transplantation can be somewhat inhibited by intrathymic injection of allogenic MHC antigen.
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Objective To study the influences of risk factors on the 2-week mortality in patients with acute cerebral hemorrhage.Methods From January 2004 to June 2007,a total of 65 patients with intracerebral hemorrhage in Beijing Red Cross Hospital were studied.Logistic regression was performed to analyze the relationship between prognostic factors and 2-week mortality.These factors included sex,age,volume of hemorrhage,location of hematoma,the highest body temperature in the first 24 h,and consciousness state on admission etc.Results The in-hospital mortality in 2 weeks was 18.5%(12/65).The volume of hemarrhage,body temperature and consciousness state were probably significant factors.Conclusion The increasing body temperature in the first 24 h and the unconsciousness may be the main risk factors which are associated with high 2-week hospital mortality in patience with intracerebral hemorrhage.
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Objective:To identify the role of transcription factors Sp1 and Sp3 in the expressional regulation of ezrin in human esophageal carcinoma cells.Methods:Esophageal carcinoma EC109 cells were transfected with expressing vectors CMV-Sp1 or CMV-Sp3,and the effect of Sp1 and Sp3 over-expression on ezrin mRNA and protein expression was determined by real time RT-PCR and Western blot analysis.Furthermore,EC109 cells were cotransfected with the ezrin promoter-directed luciferase reporter vector and control vector pRL-TK along with transcription factor expression vector.The roles of Sp1 and Sp3 in ezrin promoter activation and whether this activation occurred through the Sp1 binding site,-75/-69,were analyzed by dual-luciferase reporter assay system.Results:Over-expression of transcription factors Sp1 and Sp3 significantly increased the expression of ezrin mRNA and protein and the ezrin promoter activity in EC109 cells.Sp1 and Sp3 enhanced the promoter activity through different binding sites and only Sp1 did that through the-75/-69 site.Conclusion:Sp1 and Sp3 can regulate ezrin expression in EC109 cells.