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Objective:To investigate the efficacy of Yupingfeng powder combined with extensively hydrolyzed milk formulas in the treatment of milk protein allergy in infants and its effects on immune function, interleukin-10 (IL-10) and IL-22 levels. Methods:Eighty infants with milk protein allergy who received treatment in Hangzhou Children's Hospital from January 2020 to January 2021 were included in this study. They were randomly assigned to Yupingfeng powder and conventional treatment groups, with 40 infants per group. An additional 80 infants who concurrently received health examination in the same hospital were included in the control group. The conventional group was treated with extensively hydrolyzed milk formulas. The Yupingfeng powder group was treated with Yupingfeng powder combined with extensively hydrolyzed milk formulas. All infants were treated for 30 successive days. Clinical efficacy, serum total immunoglobulin (IgE), milk specific IgE (sIgE), peripheral blood CD 4+CD 25+ Treg, IL-10 and IL-22 levels were compared between groups. Results:Total response rate in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [92.50% (37/40) vs. 72.50% (29/40), χ2 = 5.54, P < 0.05]. Serum total IgE and milk sIgE levels in the Yupingfeng powder group were (132.93 ± 14.61) IU/L and (0.62 ± 0.14) IU/L, respectively, which were significantly lower than (150.27 ± 16.22) IU/L and (0.85 ± 0.17) IU/L in the conventional treatment group ( t = 5.02, 6.61, both P < 0.05). The expression of CD 4+CD 25+ Treg in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [(13.29 ± 1.40)% vs. (11.84 ± 1.27)%, t = 4.85, P < 0.05). CD 4+CD 25+ Treg expression in the Yupingfeng powder group was not significantly different from that in the control group [(13.40 ± 2.03)%, t = 0.31, P = 0.759]. IL-10 in the Yupingfeng powder group was significantly higher than that in the conventional treatment group [(34.57 ± 4.07) μg/L vs. (22.19 ± 2.15) μg/L, t = 17.01, P < 0.05]. IL-22 level in the Yupingfeng powder group was significantly lower than that in the conventional treatment group [(2.20 ± 0.42) ng/L vs. (5.28 ± 0.79) ng/L, t = 21.77, P < 0.05]. IL-10 and IL-22 levels in the Yupingfeng powder group were not different from those in the control group [(35.53 ± 3.85) μg/L, (2.13 ± 0.53) ng/L, t = 1.26, P = 0.209; t = 0.73, P = 0.468]. Conclusion:Yupingfeng powder combined with extensively hydrolyzed milk formulas is highly effective on milk protein allergy. The combined therapy can improve the immune function of infants, enhance IL-10 level, and decrease IL-22 level.
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Objective:To explore the risk factors of occurrence of colorectal adenoma after endoscopic polypectomy.Methods:From January 2014 to December 2019, at the Department of Day Surgery Centre in West China Hospital, Sichuan University, 6 430 patients with 20 351 polyps who underwent endoscopic colorectal polypectomy were retrospectively analyzed. Patients were divided into adenomas group (4 573 patients) and non-adenomas group (1 857 patients) according to whether they had at least one adenomatous polyp. According to the results of postoperative histopathology, colorectal polyps were divided into adenomatous polyp group (10 656 polyps) and non-adenomatous polyp group (9 695 polyps). The propensity score matching (PSM) method was applied, with 1∶1 matching, in the patients with adenoma group and the patients with non-adenoma group, as well as in adenomatous polyps group and non-adenomatous polyps group. A total of 1 824 pairs of patients and 7 362 pairs of colorectal polyps were successfully matched. After PSM, patients-related factors as gender (male), age (<40 and 40 to 60 years old), number of polyps (>2), obesity (body mass index ≥28 kg/m 2), melanosis, family history of colorectal cancer in first-degree relatives, polyps-related factors as the maximum diameter (6 to 10 and >10 mm), distribution (distal colon), and morphological classification (sessile and flat polyps) were included in the analysis of risk factors of colorectal adenoma. Univariate analysis and multivariate logistic regression were used for statistical analysis. Results:Among 6 430 patients with colorectal polyps, the detection rate of adenoma was 71.12% (4 573/6 430). After PSM, the results of univariate analysis showed that obesity, family history of colorectal cancer in first-degree relatives, the maximum diameter of polyps >10 mm were all correlated with the occurrence of adenoma (odds ratio ( OR)=1.483, 1.426 and 1.503, 95% confidence interval ( CI)1.063 to 2.067, 1.015 to 2.004, 1.198 to 1.887, all P<0.05). The results of multivariate logistic regression analysis indicated that obesity, family history of colorectal cancer in first-degree relatives, the maximum diameter of polyps >10 mm, sessile or flat polyps in morphological classification were independent risk factors of the occurence of colorectal adenomas ( OR=1.425, 1.411, 1.629, 1.165 and 1.151, 95% CI1.019 to 1.994, 1.001 to 1.988, 1.290 to 2.058, 1.030 to 1.316 and 1.012 to 1.310, all P < 0.05). Conclusions:Obesity, family history of colorectal cancer in first-degree relatives, maximum diameter of polyps >10 mm, sessile polyps or flat polyps were the independent risk factors of the occurrence of colorectal adenomas.
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Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.
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BACKGROUND:Mesenchymal stem cel s are multipotent progenitor cel s that can be isolated from the bone marrow, adipose tissue, umbilical cord blood and so on. Mesenchymal stem cel s possess strong immunosuppressive effects on both innate and adaptive immunity. OBJECTIVE:To summarize the immunomodulatory properties of mesenchymal stem cel s for T lymphocytes, B lymphocytes, natural kil er cel s and dentritic cel s and prospect its therapeutic implication. METHODS:PubMed and CNKI databases were searched for relevant literatures published from 2000 to 2015. The key words were“mesenchymal stem cel s, immunomodulator, T cel s, B cel s, NK cel s, dendriticc cel s”in Chinese and English, respectively. Then, 61 papers were included and further analyzed. RESULTS AND CONCLUSION:The immunomodulatory effects of MSCs on lymphocytes, B lymphocytes, natural kil er cel s and dentritic cel s were elicited by cel-cel contact and soluble cytokines. But mesenchymal stem cel s from different sources hold different immunomodulatory effect on immune cel s. When we use mesenchymal stem cel s in clinic, some important factors, such as isolation methods, cel sources, colonization sites, should be taken into account. In order to ensure the clinical safety and effectiveness, there are stil many problems to be further studied before mesenchymal stem cel s can be widely used in clinic.
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[ ABSTRACT] AIM:To investigate the effect of edaravone ( ED) on the expression of caspase-3 and caspase-12 in the juvenile rat hippocampus after status convulsion ( SC) .METHODS: Juvenile male Sprague-Dawley rats were ran-domly divided into normal saline ( NS) control group, SC group and ED treatment group.The rats in each group were fur-ther divided into 5 subgroups according to different time points.The rats in SC group were kindled into epilepsy by lithium-pilocarpine chemical method.The protein expression of caspase-3 and caspase-12 was determined by immunohistochemistry methods.The mRNA expression of caspase-3 and caspase-12 was detected by RT-PCR.RESULTS:(1) The IA value of caspase-3 positive cells in 24~72 h SC group increased compared with NS group.With ED intervention, the IA value of caspase-3 positive cells decreased as compared with 48~72 h SC group.The results of RT-PCR showed that the mRNA ex-pression of caspase-3 was similar to the changes of protein.( 2 ) The results of immunohistochemistry showed that the IA value of caspase-12 positive cells in 12~72 h SC group increased compared with NS group.With ED intervention, the IA value of caspase-12 positive cells decreased as compared with 24~72 h SC group.The results of RT-PCR showed that the mRNA expression of caspase-12 was similar to the changes of protein.( 3 ) In ED group, Ⅴ grade convulsion was lower than that in SC group, and the latent period of seizures in ED group was significantly longer than that in SC group.CON-CLUSION:Edaravone inhibits the expression of caspase-3 and caspase-12 in pilocarpine-induced seizures in rat hippo-campus, suggesting that edaravone has protective effect against the damage caused by status convulsion.
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BACKGROUND:The umbilical cord blood is rich of mesenchymal stem cells, which can be used as a new source of seed cells in tissue engineering. OBJECTIVE:To compare two methods for culturing, expanding and purifying the mesenchymal stem cells in vitro isolated from the human umbilical cord blood. METHODS:The ful-term birth cord blood of 40 cases was col ected under sterile conditions with heparin anticoagulation. Ficol density gradient centrifugation was used to isolate the mononuclear cells from the umbilical cord blood. The cases were randomly divided into two groups according to the different culture media. Twenty cases of umbilical cord blood were cultured in MesenGro human mesenchymal stem cells culture medium (group A), and the remaining 20 cases of umbilical cord blood were cultured in Dulbecco’s modified Eagle’s medium (group B). The occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and the number of primary cells in the two groups were compared. Cells which grew well were selected to detect the surface markers by flow cytometry. RESULTS AND CONCLUSION: The mean occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and number of primary cells in group A were better than those in group B (P < 0.01). The strong expression of the surface markers of mesenchymal stem cells (CD73 and CD105) was found by flow cytometry, of which the positive rate was 99.1%. No expression of the surface markers of hematopoietic stem cells (CD45 and CD34) was seen, of which the negative rate was 99.3%. The number, morphology, growth rate and culture time of umbilical cord blood mesenchymal stem cells cultured in MesenGro human mesenchymal stem cells culture medium were better than those cultured in Dulbecco’s modified Eagle’s medium. Cells cultured in MesenGro human mesenchymal stem cell culture medium can better express surface markers of mesenchymal stem cells.
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Objective To explore the relationship between pituitary tumor transforming gene(PTIG) and Ki67 expression and their significance in multiple adolescent breast fibroadenomas (fibroadenomatosis)and single persons(fibroadenomas) and their clinical value.Methods A total of 53 fibroadenomatosis and 160 fibroadenomas specimens in our hospital were collected.All the samples were fixed with 10％ formalin and then embedded with paraffin.The protein expression of PTTG and Ki67 was detected by immunohistochemical staining.The relation between expression of PTTG and Ki67 in fibroadenomatosis was analyzed.Results The positive rate of PTTG and Ki67 was higher in fibroadenomatosis than in fibroadenomas (P ＜ 0.005).Conclusions High expression of PTTG and Ki67 in fibroadenomatosis leads to the proliferation of tumor cell and upregulates tumor angiogenesis.High expression of PTTG and Ki67 may play important roles in tumorigenesis and pathological process of fibroadenomatosis.
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Objective To investigate the regulatory effects of miR-146a on inflammation factors production in C ryptococcus neoformans treated THP-1 cells.Methods Cryptcoo ccus neoformans strains were heat killed .Fluorescence quantitative RTP-CR and ELISA were used to detect the levels of IL -1βand TNF-αin THP-1 cells treated with heat-killed Cryptococcus neoformans.In addition, the production of IL-1βand TNF-αwere analyzed before and after Dectin-1or TLR 4 blocked .THP-1 cell lines that were respectively transfected with miR-146 a mimics and inhibitors were constructed and the production of IL-1βand TNF-αin these cell lines were analyzed after interfered with Cryptococcus neoformans.Results With the interference of heat-killed Cryptococcus neoformans, the expression of miR-146a was up-regulated in THP-1 cells, but was down-regulated after Dectin-1 or TLR4 blocked.The expressions of IL-1βand TNF-αinduced by heat-killed Cryptococcus neoformans were enhanced in miR-146a mimics transfected THP-1 cells, but was inhibi-ted in inhibitors transfected THP-1 cells.Conclusion Heat-killed Cryptococcus neoformans could up-regu-late miR-146a expression in THP-1 cells via Dectin-1 and TLR.miR-146a could negatively regulate the ex-pressions of IL-1βand TNF-αinduced by Cryptococcus neoformans.
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Objective To evaluate the diagnostic performance of plasma miR-499 in acute myocardial infarction (AMI) diagnosis.Methods Diagnostic accuracy test.The suspected AMI patients,who with chest pain for more than half an hour and been admitted in the Second People's Hospital of Wuxi and First People's Hospital of chuzhou during October 2010 and July 2011,were consecutively and prospectively enrolled in the present study.Sixty apparently healthy individuals were designed as healthy control.The plasma samples of the suspected AMI patients were collected within two hours after admission.The plasma miR-499 was determined by real time polymerase chain reaction (RT-PCR).The diagnostic performance of plasma miR-499 for AMI was estimated by receiver operating characteristic (ROC) curve analysis.The association between plasma miR-499 and AMI was analyzed by multivariable logistic model.The plasma miR-499 was determined and explained in blind fashion.Results Two hundred and nine suspected AMI patients,including 131 confirmed AMI patients (46 STEMI and 85 NSTEMI) and 78 AMI free patients were enrolled in the present study.The delta cycle threshold (ΔCT) was 1.01 ± 3.34 for AMI patients,-2.76 ± 2.90 for non-AMI patients and-3.79 ± 2.21 for healthy controls.The differences had statistical significance (x2 =96.77,P ＜ 0.01).The area under curve (AUC) of plasma miR-499 was 0.80 (95％ C I:0.74-0.86),lower than that of cardiac troponin Ⅰ (AUC =0.90,95％ CI:0.86-0.94) on admission (P ＜0.01).At the optimal cut-off of 0.18,the diagnostic sensitivity and specificity were 0.69 (95％ CI:0.61-0.77) and 0.77 (95％ CI:0.66-0.86),respectively.The coefficient of correlation between plasma miR-499 and cTnI was 0.72 (P ＜0.01).The odds ratio (OR) of plasma miR-499 ＞0.18 for AMI was 2.59 (95％ CI:1.10-6.07),after adjusted cTnI.Conclusions Plasma miR-499 is a useful biomarker for AMI diagnosis.It can provide additional diagnostic information beyond cTnI.Combination utility of plasma miR-499 and cTnI may improve the diagnostic accuracy for AMI.
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Objective To study the mechanism of mesenchymal stem cells immunosupression lym- phocyte proliferation via B7-H1/PD-1 pathway upregnlated by IFN-γ. Methods Bone marrow derived mes-enchymal stem cells (MSC) were isolated and purified by repeat adherent passage and detected them multi-potential differentiation in conditioned culture medium. Then MSC were cocuhured with lymphocyte prolifera-tion and assayed the level using γ H-thymidine incorporation. Meanwhile, ELISA measured IFN-γ, TGF-β, TNF-α and IL-10 in the cocuhured supernaatant and analyzed variation of B7-H1 molecular profile in MSC by flow cytometry. At last siRNA technology was deploied to interfere B7-H1 expression and analyzed MSC im-munosuppression on lymphocyte proliferation. Results In vitro the isolated MSC become homogeneous spi-die-shaped adherent cells after five passages, and in conditioned culture medium they could differentiate into adipocytes, osteocytes and chondrocytes. In the eocuhure of MSC with mixed lymphocyte, lymphocyte prolif- eration stimulated by Con A or by anti-CD3/CD28 antibody. The cpm value of the proliferation detected by 3H thymidine incorporation showed MSC suppressed the proliferation significantly (P = 0. 0167, 0. 0081,<0.0001 ) and the suppressive potential in a dose-dependent fashion. In the coeuhured supernatant cyto-kine IFN-T and TNF-α were detected in high concentration, but TGF-β, IL-10 were undetected. Simultane- ously MSC in the coeuhure upregulated B7-H1 expression from basic expression 7% to higher than 70% ( P < 0.05 ). After interfere B7-H1 expression in MSC by specific siRNA, we detected lymphocyte proliferation and got higher cpm by 3H thymidine incorporation ( P < 0. 05 ). Conclusion MSC upregulated B7-H1 mo-lecular expression upon the stimuli of IFN-γ, and through the B7-H1/PD-1 pathway mediated immunosu-pression on lymphocyte proliferation.
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Objective To explore a venipuncture method that can reduce the pain, improve the suc-cess rate of puncture as well as reduce vascular injury. Methods Self-controlled method was applied in 200 patients, needle tips with 20～25°,31～40°,41～50°, 51～60°, > 60° angles on the top of the blood ves-sels,the inclined plane of the needle parallel to the longitudinal axis of the blood vessel (that was,the in-clined plane to the left or right) or traditional method with upward of needle tips were adopted, the observa-tion group and the control group were divided according to different methods, pain and success rate of puncture were compared. Results There were differences between the two groups,results of the observa-tion group were better than those of the control group (except when > 60°, success rate showed no differ-ence). Conclusions Venipuncture with needle tip parallel to the longitudinal axis of the blood vessels can significantly reduce pain in patients, improve the success rate of puncture and reduce the vascular in-jury, which were superior to the traditional venipuncture method. Whatever method is chosen, 51～60° is the best angie into the vessel.
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Objective To observe the effects of in vitro isolated Schwann cells co-cultured with chemically acellular nerve allografts on improving repair of large facial nerve defects. Methods A total of 30 Wistar rats were equally randomized into three groups, ie, experimental group, allograft group and autograft group. Nerve defect of 12 mm in length was made in the left inferior buccal branch of facial nerve and repaired with acellular nerve allograft implanted with Schwann cells, acellular nerve allograft and fresh tibial nerve autograft respectively. At the 5th month postoperatively, the function and morpholo-gy of the regenerated nerves were observed by electrophysiological method, methylene blue staining and transmission electron microscope. Results In experimental group, the recovery rate (operation side/normal side) of amplitude of nerve-muscle action potential was (35.8±2.5)%, the lantency recovery rate (normal side/operation side) (65.8±2.9)%, the number of the regenerated axon 1 570±188 and the myelin thickness (0.383±0.031) μm. The results in the experimental group were significantly supe-rior to those in the acellular nerve allograft group (P < 0.05), with similar results to fresh nerve autograft group (P > 0.05). Conclusion Transplantation of Schwarm cells in acellular nerve allograft can im-prove repair of large facial nerve defects.
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Objective To explore the feasibility and superiority of the simplified partial pressurized tourniquet used in bleeding or injured limbs in battlefield.Methods The tourniquet was composed of the disposable infusion bag,bulb for blood pressure apparatus,pressure gauge and heparin cap.The disposable infusion bag wrapped around the bottom of the cravat took the places of the balloon and the harness.The pressure on the upper and lower limbs were 180mmHg and 200mmHg respectively.Results The low-cost tourniquet,with easily adjustable pressure,was easy to operate and carry.It was efficient in hemostasis and had a low rate for complications.Conclusion The tourniquet can be applied to self and buddy aid,and thus the incidence of shock can be decreased.