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Objective To analyze the epidemiological characteristics of AR and its correlation with serum IFN-γ and IL-4 levels in Bazhong City, and to provide theoretical basis for clinical prevention and treatment of AR. Methods Adopt the method of multistage stratified random survey from January 2019 to January 2020 bazhong 3 armour hospital otolaryngology seeing a doctor , with face to face questionnaire survey form the people generally, AR number of statistics, the main clinical symptoms and related symptoms, AR merger disease situation, according to the illness severity was divided into mild and moderately severe group, Five mL of elbow venous blood was extracted from AR patients, and 22 kinds of allergens were determined by using the allergen-specific IgE antibody detection kit (western blot). Serum IFN-γ and IL-4 levels were determined by enzyme-linked immunosorbent assay. Pearson correlation analysis was performed on serum IFN-γ and IL-4 levels and severity of allergic rhinitis. Results Among 1 243 patients who completed the questionnaire, the prevalence of AR was 275 (22.12%). There was significant difference in the prevalence of AR among different age groups (χ2=6.809 , P2=7.914 , P<0.0) and the prevalence of AR in workers was the highest (26.48%). Among 275 AR patients in Bazhong City, 153 cases (55.64%) had seasonal allergic rhinitis and 122 cases (44.36%) had perennial allergic rhinitis. The main clinical symptoms of AR patients were sneezing in 234 cases (85.09%) and nasal obstruction in 197 cases (71.64%). AR combined with bronchial asthma in 59 cases (21.45%), conjunctivitis in 28 cases (10.18%); the peak of AR incidence was mainly in July. Among 22 allergens, dust mite was the most common allergen in 139 cases (50.55%), followed by penicillin in 56 cases (20.36%). The main inducing factors were pollen 112 cases (40.73%) and cold 98 cases (35.64%). According to the severity of the disease, they were divided into mild group (n=178) and moderate to severe group (n=97). The serum IL-4 level in moderate and severe groups was significantly higher than that in mild group (P<0.05). The serum IFN-γ level in moderate and severe groups was significantly lower than that in mild group (P<0.05). Pearson correlation analysis showed that il-4 level was positively correlated with disease severity (r=0.492, P<0.05). IFN-γ was negatively correlated with the severity of the disease (r=-0.459, P<0.05). Conclusion The prevalence of AR is high in Bazhong city, and the main clinical symptom is sneezing. Among the complications, bronchial asthma is the most common. Colds and weather changes are the main factors causing AR.
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AIM:To observe the clinical effect of ranibizumab combined with 577nm micropulse laser in the treatment of severe diabetic macular edema(DME). METHODS:There were 52 eyes of 52 patients diagnosed with severe DME who admitted to the People's Hospital of Guangxi Zhuang Autonomous Region from June 2016 to September 2019. The patients were randomly divided into the observation group(26 patients with 26 eyes, treated with ranibizumab combined with 577nm micropulse laser)and the control group(26 patients with 26 eyes, treated with ranibizumab alone). Patients in both groups received intravitreal injection of ranibizumab with “3+PRN” regimen. Followed up at 9mo after treatment to observe the central macular thickness(CMT), the best corrected visual acuity(BCVA)and the times of intravitreal injection of ranibizumab in the two groups.RESULTS:Compared with before treatment, the CMT and BCVA of the two groups were significantly improved at each time point after treatment(all P<0.001), but there was no difference between the two groups(P>0.05). During the follow-up period, the times of vitreous injection of ranibizumabin the observation group was significantly less than that in the control group(5.88±1.24 times vs 7.12±1.24 times, P=0.001). CONCLUSION:Both ranibizumab combined with 577nm micropulse laser and ranibizumab alone are effective in reducing edema and improving vision in patients with severe DME, but the combination therapy reduces the times of injection.
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OBJECTIVE@#To explore the advantages of self made minimally invasive hook assisted transforaminal lumbar interbody fusion (TLIF) via modified bilateral Wiltse approach in the treatment of lumbar degenerative diseases.@*METHODS@#The clinical data of 140 patients underwent lumbar spine fusion surgery from October 2016 to October 2017 were retrospectively analyzed. Among them, 72 cases were treated by self-made minimally invasive hook-assisted TLIF via modified bilateral Wiltse approach (group A), there were 37 males and 35 females, aged (48±16) years old;68 cases were treated by TLIF via traditional posterior median approach (group B ), there were 38 males and 30 females, aged (45±15) years old. The surgical incision size, operation time, intraoperative blood loss volume, postoperative drainage volume, postoperative wound healing, and intervertebral fusion rate at the final follow-up were recorded between two groups. Visual analogue scale (VAS) and Oswestry Disability Index (ODI) were used to assess the clinical efficacy.@*RESULTS@#All the patients were followed up for 3 to 13 (8±5) months. The wound in group A healed well after operation, and 1 case in group B occurred wound necrosis after operation, and healed after debridement and suture. There were no significant differences in operation time and postoperative fusion rate between two surgical methods (@*CONCLUSION@#The self made minimally invasive hook assistedTLIF via modified bilateral Wiltse approach has the characteristics of minimally invasive, less intraoperative blood loss, less postoperative drainage, fewer complications, and more stable fusion in the treatment of lumbar degenerative desease.
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Adult , Female , Humans , Male , Middle Aged , Intervertebral Disc Degeneration/surgery , Lumbar Vertebrae/surgery , Minimally Invasive Surgical Procedures , Retrospective Studies , Spinal Fusion , Treatment OutcomeABSTRACT
Objective:To investigate the imaging features of pancreatic hypervascular tumors in computed tomography (CT) and magnetic resonance imaging (MRI) examinations.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 53 patients with pancreatic hypervascular tumors who were admitted to two medical centers, including 32 cases in the Affiliated Hospital of Medical School, Ningbo University and 21 cases in the First Affiliated Hospital of Naval Medical University, from March 2007 to February 2021 were collected. There were 21 males and 32 females, aged (48±23)years. Of the 53 patients, there were 19 cases with pancreatic neuroendocrine tumor (PNET), 9 cases with pancreatic metastasis from renal cell carcinoma (PRCC), 8 cases with solid pseudopapillary tumors of pancreas (SPTP), 7 cases with intrapancreatic accessory spleen (IPAS), 6 cases with serous cystadenoma of pancreas (SCP) and 4 cases with aneurysms. All the 53 patients underwent CT and MRI. Observation indicators: (1) imaging feature of PNET; (2) imaging feature of PRCC; (3) imaging feature of SPTP; (4) imaging feature of IPAS; (5) imaging feature of SCP; (6) imaging feature of aneurysms. Measurement data with normal distribution were represented as Mean±SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Imaging feature of PNET: of the 19 cases with PNET, there were 1 case with Von Hippel-Lindau disease (VHLD), 8 cases with multiple endocrine neoplasia type 1 (MEN1) and 10 cases with neuroendocrine tumor (NET). Of the 19 cases, 16 cases had single tumor and 3 cases had 2 tumors, 9 cases had tumor located at head of pancreas and 10 cases had tumor located at body and tail of pancreas. Morphology of tumors in the 19 cases were mostly round or elliptical, with some shallow lobes and clear boundary. There were 4 cases with cluster-like calcifications in the center of tumors and 15 cases with no cluster-like calcification in the center of tumors. The tumor diameter of 19 cases was (26.7±10.3)mm. Of the 19 cases, 1 case underwent pancreatic atrophy and segmental expansion of the main pancreatic duct and 18 cases underwent no pancreatic atrophy or segmental expansion of the main pancreatic duct, 2 cases underwent dilated bile ducts and 17 cases underwent no dilated bile ducts. The enhance-ment mode of imaging examination of PNET was wash in and wash out. (2) Imaging feature of PRCC: Of the 9 cases with PRCC, 2 cases had single tumor and 7 cases had multiple tumors. Of the 2 cases with single tumor, 1 case had tumor located at neck of pancreas and 1 case had tumor located at body and tail of pancreas. All the 7 cases with multiple tumors had tumor located at head, neck, body and tail of pancreas. Morphology of tumors in the 9 cases were round or quasi-circular, with clear boundary. The tumor diameter were (18.0±5.0)mm of the 2 cases with single tumor and 2.0-50.0 mm of the 7 cases with multiple tumors, respectively. Of the 9 cases, 2 cases underwent pancreatic ducts dilatation and 7 cases underwent no pancreatic ducts dilatation. The enhancement mode of imaging examination of PRCC was wash in and wash out. (3) Imaging feature of SPTP: all 8 cases with SPTP had single tumor, including 4 cases with tumor located at head of pancreas and 4 cases with tumor located at body and tail of pancreas. Morphology of tumors in the 8 cases were lobulated with clear boundary. Of the 8 cases, there were 2 cases with no calcifications of tumors and 6 cases with calcification of tumors, 2 cases with no cystic necrosis of tumors and 6 cases with cystic necrosis of tumors, 3 cases with no bleeding in the tumors and 5 cases with bleeding in the tumors. The tumor diameter of 8 cases was (51.6±11.8)mm. All the 8 cases were negative for pancreatic ducts dilatation, but the adjacent organs were compressed and moved. The enhancement mode of imaging examination of SPTP was asymptotic enhancement. (4) Imaging feature of IPAS: all the 7 cases with IPAS had single tumor located at tail of pancreas. Morphology of tumors in the 7 cases were round or quasi-circular shape with clear boundary. Of the 7 cases, 1 case with solid-cystic and uneven density tumor was epidermoid cyst in the accessory spleen of the tail of the pancreas, and 6 cases had solid and uniform density tumors. The tumor diameter of 7 cases was (25.5±8.5)mm. All the 7 cases were negative for pancreatic ducts dilatation and the surrounding structures of pancreatic ducts were clear. The enhancement mode of imaging examination of IPAS was asymptotic enhancement. (5) Imaging feature of SCP: all 6 cases with SCP had single tumor, including 1 case with tumor located at neck of pancreas and 5 cases with tumor located at body and tail of pancreas. Morphology of tumors in the 6 cases were round or quasi-circular, with clear boundary. Of the 6 cases, 2 cases had cystic tumors and 4 cases had solid tumors. The tumor diameter of 6 cases was (35.5±15.4)mm. Of the 6 cases, 2 cases were positive for pancreatic ducts dilatation and 4 cases were negative for pancreatic ducts dilatation. The enhancement mode of imaging examination of SCP was wash in and wash out. (6) Imaging feature of aneurysms: all the 4 cases with aneurysms had single tumor, including 1 case with tumor located at body of pancreas and 3 cases with tumor located at tail of pancreas. One case with tumor located at body of pancreas was superior duodenal aneurysm and 3 cases with tumor located at tail of pancreas were splenic aneurysms. Morphology of tumors in the 4 cases were round, with clear boundary. Of the 4 cases, 1 case was negative for tumor marginal calcification and 3 cases were positive for tumor marginal calcification. The tumor diameter of 4 cases was (11.3±2.5)mm. All the 4 cases were negative for pancreatic ducts dilatation. The enhance-ment mode of imaging examination of aneurysms was wash in and wash out.Conclusions:The imaging features of pancreatic hypervascular tumors in CT and MRI examinations show diversity. The enhancement mode of imaging examination of PNET, PRCC, SCP and aneurysms is wash in and wash out. The enhancement mode of imaging examination of SPTP and IPAS is asymptotic enhancement.
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Objective To investigate the resistances of larva and adult of Aedes albopictus to common insecticides in a county of Jining City, after the dengue fever outbreak in 2017, and to provide scientific support for rational use of insecticides. Methods The insecticide resistance was tested with exposure tubes for adult mosquitoes and dipping method for larvae.After collecting the larvae and breeding for one generation, the resistances of larvae were determined with the late third-instar or early fourth-instar larvae, and the resistance of adults was tested with the female mosquitoes at 3-5 days post eclosion. Results The insecticides mainly used in the county of Jining City in 2018 were pyrethroids such as permethrin, β-cyhalothrin, α-cypermethrin, and organic phosphorus pesticides such as phoxim.The median lethal concentration (LC50) values in the larva to temephos, deltamethrin, permethrin, beta-cypermethrin were 0.006 mg/L(RR=3.75), 0.021 mg/L(RR=52.50)、0.209 mg/L(RR=104.50)、0.016 mg/L (RR=17.78), with the lowest resistance to temephos and the highest to permethrin.Adult mosquitoes were sensitive to permethrin, with a knockdown rate of 97.62% and a mortality rate of 100%.When exposed to propoxur and malathion, the knockdown rates were 97.06% and 91.89%, the mortality rates were 97.06% and 93.24%.Aedes albopictus larvae were resistant to deltamethrin, α-cypermethrin, β-cyhalothrin, β-cypermethrin, with the knockdown rates of 88.00%, 95.31%, 91.04%, 97.10%, and the mortality rates were all less than 80.00%. Conclusion The larvae and adult mosquitoes of Aedes albopictus in the county of Jining City have developed different degrees of resistance to commonly used insecticides.The resistance should be under regular monitoring and the use of insecticides should be scientific and rational to delay the resistance production.
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Objective To investigate microRNAs differential expression and polarization of human macrophages in Toxoplasma gondii infection. Methods The microRNAs differential expression of human macrophages in T. gondii infection was analyzed by microarray, and further validated by qRT-PCR. pEGFP-miR-155 was transfected into THP-1 cells by Lipofectamine M2000 and the transfection ratio was detected by flow cytometry. Flow cytometry was used to detect CD86 molecular on the macrophages. qRT-PCR was used to detect iNOS and IL12 mRNA expression. NO and IL12 expression were then evaluated by using ELISA. Results The miR-155 up-regulated more than 4-fold in T. gondii infected macrophages and enhanced as well as post-infection prolong. pEGFP-miR-155 transfection ratio was 82.6%.compared to cells cultured with T. gondii, pEGFP-miR-155 and miR-155-inhibitor, T. gondii and pEGFP-miR-155 inducement enhanced expression of CD86. Additionally, iNOS and IL12 mRNA were enhanced by qRT-PCR (P<0.05). NO and IL12 expression were increased by ELISA. Conclusion T. gondii infection up-regulates the host miR-155 expression to modulate macrophages polarization to M1.
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Based on the electric field shielding and insulation technology, the single electrode dielectric barrier discharge ion source ( DBDI ) has the characteristics of uniform discharging, stability, and high ionization energy, and thus can be used to detect various samples with a large range of polarity. However, the ionization energy is too high to reduce the background signal noise, and thus affects the detection effectiveness to low polarity and low boiling point samples. To enhance the detection effectiveness to these samples, a method of eliminating electrons of single electrode DBDI by using external metal electrodes was developed in this study. Then, the single electrode DBDI was improved by an external needle electrode and an external metal net, respectively. The mechanism of those external metal electrodes was discussed, and the experimental studies were carried out. The results showed that the external metal net technology had an advantage in improving signal to noise ratio ( SNR ) , and the enhancement of SNR for the detection of isoprocarb, perfluorooctanoic acid and SudanⅢwas about 5-6 times. Based on the technology, a method for determination of Sudan Ⅲ in chili powder was developed. The recoveries, RSD and LOD were 83. 7%-94. 6% , 5. 6%-9. 0% and 23 mg/kg, respectively. The external metal electrode technique has broadened the detection range of single electrode DBDI to the field of low polarity, low boiling point and complex samples.
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Dielectric barrier discharge ion source-mass spectrometry ( DBDI-MS) can achieve in-situ and efficient detection of samples without tedious pretreatment, and has become a popular analytical method. To improve the detection sensitivity, a novel mass spectrometric method based on gold nanoparticles ( AuNPs)-assisted paper-based injection mode and DBDI-MS technology was developed for the fast determination of four veterinary drugs. The detection signal strength was improved by 8-31 times compared to the veterinary drugs which were tested without AuNPs-assisted paper based injection. Under the optimal conditions, the experimental data of four veterinary drugs showed a good linear relationship between the signal and the concentration in the respective range, with correlation coefficient ( r ) of 0. 951-0. 996. Moreover, the recoveries were between 79. 4% and 96. 5% at three spiked levels. This method has many distinct advantages, such as low detected limitation, low cost and high efficiency, and is also very suitable for fast on-site detecting, showing great potential in feed quality control.
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The TLC method was established for identification of Holotricha diomphalia larvae and the HPLC method was used to determine the content of inosine and guanosine in H. diomphalia larvae. The HPLC analysis was performed on a Waters HSS T3(4.6 mm×250 mm, 5 μm) column of with mobile phase consisting of acetonitrile (A) and 0.08% trifluoroacetic acid (B) in gradient elution. The detection wavelength was 260 nm. The flow rate was 1.0 mL·min⁻¹. The column temperature was 30 °C. As a result, TLC identification method had a good reproducibility and highly specificity. The linear equations of inosine and guanosine were in good linear range (r>0.999 8). The average recovery of inosine and guanosine was 96.53% (RSD=1.6%), 99.71% (RSD=2.7%). The method is simple, accurate and reproducible, which can provide a basis for quality standard improvement H. diomphalia larvae.
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Objective To investigate microRNAs differential expression and polarization of human macrophages in Toxoplasma gondii infection. Methods The microRNAs differential expression of human macrophages in T. gondii infection was analyzed by microarray, and further validated by qRT-PCR. pEGFP-miR-155 was transfected into THP-1 cells by Lipofectamine M2000 and the transfection ratio was detected by flow cytometry. Flow cytometry was used to detect CD86 molecular on the macrophages. qRT-PCR was used to detect iNOS and IL12 mRNA expression. NO and IL12 expression were then evaluated by using ELISA. Results The miR-155 up-regulated more than 4-fold in T. gondii infected macrophages and enhanced as well as post-infection prolong. pEGFP-miR-155 transfection ratio was 82.6%.compared to cells cultured with T. gondii, pEGFP-miR-155 and miR-155-inhibitor, T. gondii and pEGFP-miR-155 inducement enhanced expression of CD86. Additionally, iNOS and IL12 mRNA were enhanced by qRT-PCR (P<0.05). NO and IL12 expression were increased by ELISA. Conclusion T. gondii infection up-regulates the host miR-155 expression to modulate macrophages polarization to M1.
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BACKGROUND: Our study aimed to investigate the roles of autophagy against high glucose induced response in retinal pigment epithelium (ARPE-19 cells). METHODS: The morphological changes and reactive oxygen species (ROS) generation in ARPE-19 cells under high glucose treatment were respectively detected using the transmission electron microscopy and flow cytometry. The expression levels of Parkin, PINK1, BNIP3L, LC3-I and LC3-II in ARPE-19 cells received high glucose treatment were measured by western blot after pretreatment of carbonyl cyanide m-chlorophenylhydrazone (CCCP), 3-methyladenine (3-MA), N-acetyl cysteine (NAC) or cyclosporin A (CsA) followed by high glucose treatment. RESULTS: ARPE-19 cells subjected to high glucose stress showed an obvious reduction in the LC3-I expression and significant increase in the number of autophagosomes, in the intracellular ROS level, and in the expression levels of Parkin, PINK1, BNIP3L and LC3-II (p < 0.05). Pretreatment with CCCP significantly reduced the LC3-I expression and increased the expression levels of Parkin, PINK1, BNIP3L and LC3-II (p < 0.05). ARPE-19 cells pretreated with CsA under high glucose stress showed markedly down-regulated expressions of Parkin, PINK1 and BNIP3L compared with the cells treated with high glucose (p < 0.05). Pretreatment of ARPE-19 cells with NAC or 3-MA under high glucose stress resulted in a marked reduction in the expression levels of PINK1, BNIP3L and LC3-II (p < 0.05). Meanwhile, the expression level of Parkin in the ARPE-19 cells pretreated with NAC under high glucose stress was comparable with that in the control cells. CONCLUSION: Autophagy might have protective roles against high glucose induced injury in ARPE19 cells via regulating PINK1/Parkin pathway and BNIP3L.
Subject(s)
Humans , Protein Kinases/drug effects , Autophagy/drug effects , Proto-Oncogene Proteins/drug effects , Tumor Suppressor Proteins/drug effects , Ubiquitin-Protein Ligases/drug effects , Retinal Pigment Epithelium/drug effects , Glucose/pharmacology , Membrane Proteins/drug effects , Protein Kinases/metabolism , Autophagy/physiology , Signal Transduction/physiology , Cell Line , Proto-Oncogene Proteins/metabolism , Reactive Oxygen Species/metabolism , Tumor Suppressor Proteins/metabolism , Ubiquitin-Protein Ligases/metabolism , Microscopy, Electron, Transmission , Retinal Pigment Epithelium/cytology , Flow Cytometry , Membrane Proteins/metabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes of hepcidin and ferropotin 1 expression in murine model of iron overload.</p><p><b>METHODS</b>The murine model of iron overload was established, C57BL/6 mice were injected with iron dextran intraperitoneally (10 mg) every 3 days for 4 weeks. Blood routine, serum ferritin and pathological sections were tested at the appointed time-point respectively (before iron injection, 2 weeks and 4 weeks after treatment of iron injection). The serum hepcidin was assayed by enzyme-linked immunosorbent method. The expression of ferroportin 1 in bone marrow cells was detected by RT-PCR and Western blot, respectively. The labile iron pool of bone marrow cells was measured by flow cytometry.</p><p><b>RESULTS</b>The absolute number and percentage of reticulocytes in the iron-overloaded mice were significantly decreased along with the increase of iron injection times (r=-0.938, r=-0.947), while no significant change was found in the number of white blood cells, hemoglobin level and platelet count. The level of serum ferritin was increased along with increase of iron injection time (r=0.894). Iron overload was found in pathological sections of different organs. Furthermore, serum hepcidin was increased along with increase of iron injection time (r=0.957). RT-PCR and Western blot analyses showed that the expressions of ferroportin 1 at mRNA and protein level were increased in the murine model of iron overload (P<0.05). Labile iron pool in bone marrow cells was also found to be increased in the murine model of iron overload(P<0.05).</p><p><b>CONCLUSION</b>The expressions of hepcidin and ferroportin 1 are increase in a murine model of iron overload, which may be contributed to the suppression effect on erythropoiesis in bone marrow.</p>
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Shanxi, a traditional production area to produce genuine Astragali Radix of high quality, has experienced major changes in the pattern of resources. This area once accounted for half of Astragali Radix industry, but now only serves as the largest supply area of traditional wild Astragali Radix. Furthermore, the strategic position of Shanxi Astragali Radix industry will become more prominent and more important to economic and social development in face of the diversity of market demands, especially for the strong demands of high-end Astragali Radix. In addition, Astragalus industry involves the simultaneous development of the first, second and tertiary industries in many areas, and it is typical and representative in the traditional Chinese medicine industry development. However, the application and industrial development of Shanxi Astragali Radix have been restricted due to the problems such as blind promotion of transplanting cultivation technology, and lack of science and technology including efficacy investigation, safety evaluation, standardization and controllability studies. Therefore, we would analyze the production history, resource structure, the current situation and progress of industry development, scientific research foundation and existing problem in this paper, and put forward countermeasures for development and technical innovation in order to make Astragali Radix industry bigger and stronger through innovation-driven and make benefits for demos. This thought provides a reference for the exploratory development of other large varieties of Chinese medicinal materials.
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<p><b>OBJECTIVE</b>To investigate the correlation of the autophagy-associated gene Atg5 with the pathogenesis of prostate cancer.</p><p><b>METHODS</b>Using real-time fluorescent quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and immunohistochemistry, we detected the expression of Atg5 in 50 cases of prostate intraepithelial neoplasm (PIN), 69 cases of prostate cancer (PCa), and 30 cases of benign prostatic hyperplasia (BPH).</p><p><b>RESULTS</b>The expression level of Atg5 mRNA was significantly higher in PIN (5.270 ± 0.230) and PCa (5.131 ± 0.252) than in the BPH tissue (1.723 ± 0.017) (P <0.01), and so was the positive rate of the Atg5 expression in the patients of the PIN group (94%) and PCa group (88.4%) than in those of the BPH group (6.7%) (P<0.01), but with no statistically significant differences between the PIN and PCa groups (P >0.05). No significant correlation was observed between the expression of Atg5 and the Gleason score of PCa (P >0.05).</p><p><b>CONCLUSION</b>The upregulated expression of Atg5 might play a role in the tumorigenesis of prostate cancer.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Autophagy , Autophagy-Related Protein 5 , Immunohistochemistry , Microtubule-Associated Proteins , Genetics , Prostatic Hyperplasia , Genetics , Prostatic Neoplasms , Genetics , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Methods , Up-RegulationABSTRACT
The aim of this study was to investigate the effects of CD4(+)CD25(+) regulatory T cells (Treg) on allogeneic hematopoietic stem cell transplantation (HSCT) in sensitized mice so as to provide experimental evidence for clinical treatment of allogeneic HSCT rejection in sensitized recipients. The BALB/c mice were divided into 5 groups: group A - mice were sensitized with injection of splenocytes; group B - mice were sensitized with splenocytes and treated with >5×10(5) Treg on day 7 before transplantation; group C - mice were sensitized with splenocytes and treated with 5×10(5) Treg on day 13 and 7 before transplantation; group D - mice were not sensitized, but treated with equal volume of PBS as control; group E - blank control. Each group had 15 mice. On day 0 of transplantation, mice in each group were irradiated lethally with 8 Gy by linear accelerator, and the bone marrow cells of C57BL/6 labeled by fluorescence staining were intravenously injected via the tail vein. The fluorescent cells in peripheral blood and organ tissue were detected by flow cytometry on different time points for homing assessment. Survival rates and hematopoietic reconstitution were also recorded and monitored. The results showed that on 12 and 24 hours after transplantation, as compared with the sensitized group, the number of fluorescence homing cells in different tissue of the applied Treg groups increased significantly and the differences were statistically significant (P < 0.05). The mice in sensitized group and blank control group all died on the 6-13 day, whereas the median survival time of mice in applied Treg once and twice were 15 days and 16 days respectively. Comparing with sensitized group, the difference was statistically significant (P < 0.001), but there was no significant difference between these two groups applied regulatory T cell (P > 0.05). It is concluded that applying Treg can induce immune tolerance of sensitized recipient to allogeneic HSCT and inhibit immune destruction and prolong the survival time, but can not induce full immune tolerance and at last sensitized mice died of rejection of hematopoietic stem cells.
Subject(s)
Animals , Male , Mice , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Methods , Immune Tolerance , Mice, Inbred BALB C , Mice, Inbred C57BL , T-Lymphocytes, Regulatory , Allergy and Immunology , Transplantation, HomologousABSTRACT
This study was aimed to explore the effects of blocking B7/CD28 and CD40/CD154 co-stimulatory signals on immune function of sensitized mice', and provide the evidences of acquired immune tolerance for allogeneic bone marrow transplantation. The mice sensitized on 7 day before transplant were divided into 4 groups: (1)CTLA4Ig+ anti-CD154 isotype control IgG; (2)anti-CD154 +CTLA4Ig isotype control IgG; (3)CTLA4Ig and anti-CD154; (4)isotype control IgG of CTLA4Ig and anti-CD154. CTLA4Ig and anti-CD154 used in normal BALB/c mice as isotype control IgG. Each mouse in all groups received CTLA4Ig and anti-CD154 (or corresponding isotype control IgG) 500 µg respectively, and was injected via tail vein on 7 day before transplant. There were 5 mice in each group. The mice were sacrificed on day 0, then the number of CD19(+)CD69(+)B cells, CD44(high)/CD62L(high) and CD44(high)/CD62L(low)/- T cells were measured by flow cytometry. Changes of cytokines and sensitized antibody were tested by ELISA or flow cytometry. The results showed that the numbers of CD19(+)CD69(+)B cells were significantly increased in comparison with the normal group (P < 0.01) , whereas the numbers of cells were significantly decreased when blocking B7/CD28 or /and CD40/CD154 co-stimulatory signals (P < 0.01) . Blocking these 2 signals together displayed a synergistic effect (P < 0.01) . The central memory and effector T cells were defined as CD44(high)/CD62L(high) and CD44(high)/CD62L(low)/- respectively, those increased significantly after sensitized in comparison with those in normal group, whereas their numbers decreased when blocking B7/CD28 or/and CD40/CD154 co-stimulatory signals. Blocking these two signals together, displayed a synergistic effect (P < 0.01). Cytokines, IgG and IgM in all groups were not significantly different. Sensitizing antibody test showed that the fluorescence intensity of sensitized group significantly increased as compared with normal group, whereas fluorescence intensity of CTLA4Ig or/and anti-CD154 treated groups significantly decreased as compared with sensitized group (P < 0.01) . It is concluded that blocking the B7/CD28 or/and CD40/CD154 co-stimulatory signal can inhibit the cellular and humoral immune function, whereas blocking these two signals together displays a synergistic effect.
Subject(s)
Animals , Male , Mice , B7-1 Antigen , Metabolism , Bone Marrow Transplantation , CD28 Antigens , Metabolism , CD40 Antigens , Metabolism , CD40 Ligand , Metabolism , Immune Tolerance , Allergy and Immunology , Mice, Inbred BALB C , Mice, Inbred C57BL , Signal Transduction , Transplantation, HomologousABSTRACT
The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.
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Humans , Cell Membrane , Metabolism , Endosomes , Metabolism , Gene Transfer Techniques , Genetic Therapy , Genetic VectorsABSTRACT
The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.
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<p><b>BACKGROUND</b>The effects of chest compressions to hemodynamic and respiratory parameters during hands-only cardiopulmonary resuscitation (CPR) in a non-arrested patient who suddenly collapses are confusing. In this research, we investigated the effects of chest compressions in a non-arrested porcine model.</p><p><b>METHODS</b>Fourteen male domestic pigs were randomized into sham control group (SHAM group, only anesthetized and instrumented without chest compression, n = 6) or chest compression group (CC group, 2 minutes of chest compressions, n = 8). Continuous hemodynamic parameters, dynamic lung compliance (Cdyn), and blood gas analysis outcomes were recorded. Serum levels of catecholamine were measured at baseline and 2 minutes, 30 minutes, 4 hours, and 24 hours after chest compressions. Chest computed tomography (CT) was performed at 30 minutes and 24 hours. Conventional histopathology evaluation was performed.</p><p><b>RESULTS</b>After two minutes of chest compressions in the CC group, heart rate and extravascular lung water increased significantly; mean arterial pressure, stroke volume, and global ejection fraction significantly decreased. Cdyn significantly decreased to valley levels at 30 minutes and slowly recovered. Compared with the baseline, serum levels of catecholamine significantly increased at 2 minutes and rapidly decreased 24 hours later. At 30 minutes after chest compressions, chest CT showed local exudation, which was absorbed 24 hours later.</p><p><b>CONCLUSIONS</b>This research showed that 2 minutes of chest compressions causes various heart and lung tissue damage in the normal a normal porcine model. It also impacts the hemodynamic and Cdyn.</p>
Subject(s)
Animals , Male , Cardiopulmonary Resuscitation , Catecholamines , Blood , Extravascular Lung Water , Heart Massage , Heart Rate , Hemodynamics , Lung Compliance , Models, Animal , Swine , Tomography, X-Ray ComputedABSTRACT
The study was purposed to explore the effect of panel reactive antibody (PRA) serum from patients with β-thalassemia on proliferation and apoptosis of the CD34(+)cells from cord blood and its mechanism. CD34(+) cells of umbilical cord blood were incubated with different sera and complement respectively. After incubation, the samples were centrifuged and the supernatants were collected for lactate dehydrogenase (LDH) detection, and the CD34(+) cells were harvested and measured for the apoptosis by flow cytometry with Annexin V/PI. The intracellular DNA synthesis were also quantified by [(3)H]TdR incorporation using liquid scintillation counter. The results showed that concentration of LDH in PRA positive groups was higher as compared with control group, and the DNA synthesis of CD34(+) cells in PRA positive groups were inhibited. There were no differences in the percentage of cell apoptosis and necrosis among different groups. It is concluded that thalassemic serum PRA impairs the cell membrane, inhibits the DNA synthesis, which can be increased by addition of the complement, but PRA had no significant effect on apoptosis of CD34(+) cells.