Decellularized matrix transplantation has emerged as a promising therapeutic approach for repairing tissue defects, with numerous studies assessing its safety and efficacy in both animal models and clinical settings. The host immune response elicited by decellularized matrix grafts of natural biological origin plays a crucial role in determining the success of tissue repair, influenced by matrix heterogeneity and the inflammatory microenvironment of the wound. However, the specific immunologic mechanisms underlying the interaction between decellularized matrix grafts and the host immune system remain elusive. This article reviews the sources of decellularized matrices, available decellularization techniques, and residual immunogenic components. It focuses on the host immune response following decellularized matrix transplantation, with emphasis on the key mechanisms of Toll-like receptor, T-cell receptor, and TGF-β/SMAD signaling in the stages of post-transplantation immunorecognition, immunomodulation, and tissue repair, respectively. Furthermore, it highlights the innovative roles of TLR10 and miR-29a-3p in improving transplantation outcomes. An in-depth understanding of the molecular mechanisms underlying the host immune response after decellularized matrix transplantation provides new directions for the repair of tissue defects.

Objective:To summarize the clinical characteristics of patients with coronavirus disease 2019 (COVID-19) infected with Delta variant, so as to provide further references for clinical diagnosis and treatment.Methods:A real-world study was conducted to analyze the characteristics of 166 COVID-19 patients infected with Delta variant at Guangzhou Eighth People’s Hospital, Guangzhou Medical University.Results:The study enrolled 5 asymptomatic cases, 123 non-severe cases (mild and moderate type), and 38 severe cases (severe and critical type). Among these patients, 69 (41.6%) were male and 97 (58.4%) were female, with a mean age of 47.0±23.5 years. Thirty-nine cases (23.5%) had received 1 or 2 doses of inactivated vaccine. The incidence of severe COVID-19 cases was 7.7% in 2-doses vaccinated patients, which was lower than that of 11.5% in 1-dose and 26.8% in unvaccinated patients. The proportion of severe cases in 2 dose-vaccinated patients was 7.7%, which was lower than that of 11.5% in 1-dose vaccinated patients and 26.8% in unvaccinated patients, but the difference was not significant ( P>0.05). The most common clinical symptom was fever (134 cases, 83.2%), and 39.1% of cases presented with high-grade fever (≥39 °C); other symptoms were cough, sputum, fatigue, and xerostomia. The proportion of fever in severe cases was significantly higher than that of non-severe cases (97.4% vs. 76.4%, P<0.01). Similarly, the proportion of severe cases with high peak temperature (≥39 ℃) () was also higher than that of non-severe cases (65.8% vs. 30.9%, P<0.01). The median minimal Cycle threshold (Ct) values of viral nucleic acid N gene and ORFlab gene were 20.3 and 21.5, respectively, and the minimum Ct values were 11.9 and 13.5, respectively. Within 48 h of admission, 9.0% of cases presented with decreased white blood cell counts, and 52.4% with decreased lymphocyte counts. The proportions of increased C-reactive protein, serum amyloid A, interleukin 6, and interleukin 10 were 32.5%, 57.4%, 65.3%, and 35.7%, respectively. The proportions of elevated C-reactive protein, serum amyloid A and interleukin-6 in severe cases were significantly higher than those in non-severe cases ( P<0.01). Logistic regression analysis showed that older age and higher peak temperature were associated with a higher likelihood of severe cases ( OR>3, 95% CI: 2-7, P<0.01). In terms of treatment, traditional Chinese medicine (TCM) was used in 97.6% of non-severe cases and 100% in severe cases. Other treatments included respiratory and nutritional support, immunotherapy (such as neutralizing antibodies and plasma of recovered patients). The median times from admission to progression to severe cases, of fever clearance, and of nucleic acid conversion were 5 days, 6 days and 19 days, respectively. No deaths were reported within 28 days. Conclusions:The symptoms of Delta variant infection in Guangzhou are characterized by a high proportion of fever, high peak temperature, long duration of fever, high viral load, a long time to nucleic acid conversion, and a high incidence of severe cases. The severe cases exhibit a higher percentage of elderly patients, a longer duration of fever and have a higher fever rate and a higher hyperthermia rate than non-severe cases. Age and hyperthermia are independent risk factors for progression to severe disease. The combination of TCM and Western medicine can control the progression of the disease effectively.

At present, acellular matrix is an effective replacement material for the treatment of skin damage, but there are few systematic evaluation studies on its performance. The experimental group of this study used two decellularization methods to prepare the matrix: one was the acellular matrix which sterilized with peracetic acid first (0.2% PAA/4% ethanol solution) and then treated with hypertonic saline (group A), the other was 0.05% trypsin/EDTA decellularization after γ irradiation (group B); and the control group was soaked in PBS (Group C). Then physical properties and chemical composition of the three groups were detected. Hematoxylin eosin (HE) staining showed that the acellular effect of group B was good. The porosity of group A and B were both above 84.9%. In group A, the compressive modulus of elasticity was (9.94 ± 3.81) MPa, and the compressive modulus of elasticity was (12.59 ± 5.50) MPa in group B. There was no significant difference between group A or B and group C. The total content of collagen in acellular matrix of group A and B was significantly lower than that of group C (1. 662 ± 0.229) mg/g, but there was no significant difference in the ratio of collagen Ⅰ/Ⅲ between group B and group C. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that there was no significant difference in microstructure. Qualitative detection of fibronectin and elastin in each group was basically consistent with that in group C. Therefore, acellular matrix of group B had better performance as scaffold material. The experimental results show that the acellular matrix prepared by γ-ray sterilization and decellularization of 0.05% Trypsin enzyme/EDTA could be used for the construction of tissue-engineered skin. It could also provide reference for the preparation and mounting of heterogeneous dermal acellular matrix. It was also could be used for electrostatic spinning or three-dimensional printed tissue engineered skin scaffold which could provide physical and chemical parameters for it.
Acellular Dermis ; Cells, Cultured ; Extracellular Matrix ; Porosity ; Tissue Engineering ; Tissue Scaffolds

Objective: To explore the inhibitory effect of icotinib combined with cytokine induced killer (CIK) on various human lung adenocarcinoma cell lines in vitro. Methods: The inhibitory effect of icotinib alone or icotinib combined with CIK on HCC827 and A549 cells was detected by cell counting kit-8(CCK-8). The apoptosis was detected by flow cytometry via Annexin V/PI staining. The effect of icotinib on CIK phenotype was detected by flow cytometry. Results: The inhibitory rates of HCC827 cells treated with 1.5, 3, 6, 12 μmol/L icotinib were (5.64±0.05)%, (8.62±0.45)%, (14.57±0.65)% and (18.52±0.91)%, respectively. The inhibitory rates of A549 cells were (1.64±0.48)%, (2.09±0.28)%, (3.69±0.45)%, (4.41±0.58)%, respectively. At the same concentration, the inhibitory rate of HCC827 cells with icotinib treatment was significantly higher than that of A549 cells (P<0.05). When the effector/target ratio was 10∶1, 20∶1 or 40∶1, the inhibitory rates of HCC827 cells co-cultured with CIK were (15.17±2.33)%, (42.59±7.18)%, (62.59±8.95)%, respectively, and the inhibitory rates of A549 were(16.99±2.81)%, (46.31±1.89)%, (58.24±4.23)%, respectively. The inhibitory rate of HCC827 cells co-cultured with CIK was not significantly different from that of A549 cells at the same effector/target ratio (P_10∶1=0.299, P_20∶1=0.318, P_40∶1=0.366). When the effector/target ratio of CIK combined with 6 μmol/L icotinib was 10∶1, 20∶1 or 40∶1, the inhibitory rates of HCC827 cells were (37.07±3.50)%, (76.03±6.55)%, (80.34±10.69)%, respectively, and the inhibitory rates of A549 cells were(25.72±1.41)%, (52.76±3.82)%, (62.26±1.94)%, respectively. The inhibitory rates of 6 μmol/L icotinib combined with CIK were significantly higher than those of icotinib group and CIK group alone at the same effector/target ratio (P<0.05), except for the effector/target ratio at 40︰1 on A549 cells (P=0.089). Moreover, all of the combination index (CI) of combined group were <1 (P<0.05). The apoptotic rates of HCC827 and A549 cells induced by icotinib combined with CIK were significantly higher than those of icotinib group and blank control group (P<0.05), especially the proportion of late apoptotic or necrotic cells.Increasing effector/target ratio of CIK contributed to stronger inhibition(P<0.05). The expressional rate of CIK phenotype with or without icotinib treatment was not significantly different from each other(P>0.05). Conclusions EGFR mutant lung adenocarcinoma cells are more sensitive to icotinib, while the EGFR mutation status has no effect on the killing effect of CIK cells. icotinib combined with CIK has a synergistic effect on the inhibition of tumor growth, and icotinib has no any impact on the phenotype of CIK cells.

Objective Using abandoned white cells separated from preparation of blood products to cultivate NK cells in vitro, and to optimize the method of cultivation of allogeneic NK cells for clinical application.Methods Abandoned white cells separated from blood production were collected from 15 healthy donors.PBMCs were isolated from the abandoned white cells and cultured for 17 days using culture bottles as previously coated antibodies (group CD3 mAb was coated with CD3 mAb, group CD 16mAb was coated with CD16mAb, and group CD3mAb +CD16 mAb was coated with CD3 mAb and CD16 mAb).Flow cytometry was used to determine the ratio of CD3-CD56+cells, expression of activated cell surface receptors, and secretion of IFN-γ.The anti-tumor cytotoxicity against K562 and Raji cells was determined using LDH cytotoxicity assay and flow cytometry.Results After expansion for 17 days, the proportions of CD3-CD56+cells was (15.19±12.22)% in the group CD3 mAb, (83.63±10.63)% in the group CD16 mAb, (49.40± 12.64)% in the group CD3mAb +CD16 mAb, and it was (16.34±10.51)% before expansion.The total number of NK cells was more than 10 9 .The expression ratios of NK cell surface activated receptors NKp30 and NKp46 were significantly increased, while that of the NKG2D was not significantly changed.The NK cells after expansion showed high cytotoxicity activity against K562 cells, reaching up to(76.97±3.16)%when effector-cell-to-target-cell ratio (E ∶T ratio) was 40 ∶1.Conclusions NK cells can be obtained from abandoned white cells after cultivation for 17 days, with a purity up to 90% and total cell number of more than 10 9 .Their activity was reinforced, the anti-tumor cytotoxicity activity was increased, and may meet the standard of clinical therapeutic application.

Objective Using abandoned white cells separated from preparation of blood products to cultivate NK cells in vitro, and to optimize the method of cultivation of allogeneic NK cells for clinical application.Methods Abandoned white cells separated from blood production were collected from 15 healthy donors.PBMCs were isolated from the abandoned white cells and cultured for 17 days using culture bottles as previously coated antibodies (group CD3 mAb was coated with CD3 mAb, group CD 16mAb was coated with CD16mAb, and group CD3mAb +CD16 mAb was coated with CD3 mAb and CD16 mAb).Flow cytometry was used to determine the ratio of CD3-CD56+cells, expression of activated cell surface receptors, and secretion of IFN-γ.The anti-tumor cytotoxicity against K562 and Raji cells was determined using LDH cytotoxicity assay and flow cytometry.Results After expansion for 17 days, the proportions of CD3-CD56+cells was (15.19±12.22)% in the group CD3 mAb, (83.63±10.63)% in the group CD16 mAb, (49.40± 12.64)% in the group CD3mAb +CD16 mAb, and it was (16.34±10.51)% before expansion.The total number of NK cells was more than 10 9 .The expression ratios of NK cell surface activated receptors NKp30 and NKp46 were significantly increased, while that of the NKG2D was not significantly changed.The NK cells after expansion showed high cytotoxicity activity against K562 cells, reaching up to(76.97±3.16)%when effector-cell-to-target-cell ratio (E ∶T ratio) was 40 ∶1.Conclusions NK cells can be obtained from abandoned white cells after cultivation for 17 days, with a purity up to 90% and total cell number of more than 10 9 .Their activity was reinforced, the anti-tumor cytotoxicity activity was increased, and may meet the standard of clinical therapeutic application.

Objective To explore the effect of duraphat varnish on reducing orthodontic tooth enamel demineralization around brackets.Methods We Selected 30 patients aged 12 to 14 years old in orthodontic Departrnent of Shenyang Stomatological Hospital from Jan 2011 to Dec 2013 and carried out rectification scheme to pull out the first premolar after test.The full mouth dental were divided into four parts by the quadrant and the first premolars of different groups were coated with Tooth Mousse,Fluor Protector and saline (as control group),duraphat varnish (as experimental group) respectively.Every group included 30 teeth.Three months later,We observed the demineralization of the teeth.The enamel decalcification of all quarters were detected by DI-AGNOdent.Results The rate of enamel demineralization in the experimental group was 10.0％,that in the Tooth Mousse group was 13.3％,the 0.1％ Fluor Protector group 23.3％,the saline group 53.3％.There were significant statistical difference of the rate of enamel demineralization between the Duraphat varnish group and 0.1％ Fluor Protector group,and that between the Duraphat varnish group and the saline group (P ＜ 0.01).There was no statistical difference of that between the Duraphat varnish group and the Tooth Mousse group (P ＞0.05).There were no statistical difference of DIAGNOdent reading between the experimental group and the control groups before bonding(P ＞ 0.05).After bonding,one month later and three months later,there was no statistical difference of DIAGNOdent reading between the Duraphat varnish group and the Tooth Mousse group (P ＞ 0.05).There was significant statistical difference of that between the Duraphat varnish group and 0.1％ Fluor Protector group(P ＜ 0.01).Conclusion Duraphat varnish can reduce the tooth enamel demineralization more effectively than 0.1％ Fluor Protector and saline in orthodontic treatment,and also can be used for children who were wearing fixed orthodontic appliances.

Objective:To discuss the effect and mechanism of autophagy inhibitor 3-MA on arsenic trioxide inducing apoptosis of acute T-cell leukemia cell line Jurkat cells.Methods:Proliferation inhibition of Jurkat cells treated with arsenic trioxide was detected by XTT.Morphological characteristics of Jurkat cells treated with different concentrations arsenic trioxide were observed by electron mi-croscope.Microtubule-associated protein 1 light chain 3B (LC-3B) protein expression was detected by Western blot and flow cytome-try.Apoptosis rates of Jurkat cells treated with 3-MA combining arsenic trioxide were detected by flow cytometry using AnnexinV-FITC/PI double staining.Results:Arsenic trioxide inhibited the growth of Jurkat cells in a dose and time dependence.We observed different morphological characteristics of autophagy , apoptosis and necrosis accompanying more autophagosomes in Jurkat cells which were treated with arsenic trioxide 2.5,5,10 μmol/L after 24 h.LC3B mean fluorescence intensity (MFI)relative multiples were(3.1±0.2) fold,(4.6±0.31)fold,(34.2±4.5)fold with 5 μmol/L arsenic trioxide treated Jurkat cells 0,24,48 h,and the P values between each of the two groups were less than 0.05,which increased depending time consistently with the growth inhibition rates.LC-3B protein expression gradually increased treated Jurkat cells with arsenic trioxide after 24 h,48 h.The growth inhibition rate (60.6±8.3)%was significantly different treated with arsenic trioxide combining 3-methyl adenine ( 3-MA ) while it was ( 33.4 ±9.1 )% treated with arsenic trioxide alone, however, LC-3B protein expression gradually decreased.Jurkat cell apoptosis rate ( 44.96 ±3.60 )% was significantly increased treated with arsenic trioxide combining autophagy inhibitor(3-MA) while it was (2.94±0.26)% treated with arsenic trioxide alone, and this difference was statistically significant.Conclusion: 3-MA increased apoptosis rates of Jurkat cells inducing by Arsenic trioxide and it may be related with inhibition of autophagy and induction of apoptosis.

Objective To investigate nasomaxlllary complex facial soft tissue changes after the treatment with maxillary protraction appliance with skeletal Class Ⅲ malocclusion with a retruded maxilla.Methods Thirty growing subjects with skeletal Class Ⅲ malocclusions with maxillary retrognathism were selected and treated by facial mask(male 15,female 15,with an average age of 10.5).They were given a maxillary protraction treatment with face mask for 6-8 months.Cephalometric measurements about nasomaxillary complex soft tissue changes were analyzed to draw the statistic conclusion.Results After maxillary protraction treatment,PraY,nasofrontal angle,As-Y,UL-Y,UL-E,S-Ns-Sn increased (P ＜ 0.01) ; M-Y increased (P ＜ 0.05) ; LL-E,PosY,nasolabial angle decreased (P ＜ 0.05).There were no significant differences in the Ns-Y and columella-tip angle.Conclusion After maxillary protraction treatment,nasomaxillary complex area becomes more marked.Both the nasomaxillary complex soft tissue and lower facial profile are dramatically improved.The combining effect of these two changes results in a more harmony profile.

Objective To compare the efficacy of zillah risperidone and risperidone in the treatment of schizophrenia. Methods In our hospital from January 2010 to January 2014,selected 60 cases schizophrenic patients hospitalized as observed object,60 cases hospitalized patients with schizophrenia during as the observed object, all patients were ran-domly divided into group A (ziprasidone) and group B (risperidone), each group of 30 patients, before and after treatment efficacy PANSS score changes and two groups of patients after treatment, adverse reactions were compared between two groups. Results After 4,8 weeks, the PANSS total score,PANSS positive symptoms, PANSS negative symptoms of A group and B group were significantly lower than before treatment,the difference was statistically signifi-cant(P<0.05),PANSS scores of A group after treatment 4,8 weeks were significantly lower than in group B,the differ-ence was statistically significant (P<0.05),but after treatment 4,8 weeks,PANSS positive symptoms, PANSS negative symptoms of group A and group B had no significant significance. The total effective rate of A group was 86.7%,was higher than that in group B (83.3%),but the difference was not statistically significant between the two groups (P>0.05). After treatment for 6 weeks,the incidence of adverse reactions of A group was 20%,group B was 40%,the inci-dence of adverse reactions in group A was significantly lower than group B,the difference was statistically significant(P<0.05). Conclusion Ziprasidone and risperidone in the treatment of schizophrenia have better efficacy, but com-pared to risperidone, ziprasidone has advantages,with fewer adverse reactions.