BACKGROUND:Compared with traditional two-dimensional culture,three-dimensional microtissue culture can show greater advantages.However,more favorable cultivation methods in three-dimensional culture still need to be further explored. OBJECTIVE:To evaluate the cell behavior of microtissue and its ability to promote cartilage formation under two three-dimensional culture methods. METHODS:Cartilage-derived microcarriers were prepared by chemical decellularization and tissue crushing.DNA quantification and nuclear staining were used to verify the success of decellularization,and histological staining was used to observe the matrix retention before and after decellularization.The microcarriers were characterized by scanning electron microscopy and CCK-8 assay.Cartilage-derived microtissues were constructed by combining cartilage-derived microcarriers with human adipose mesenchymal stem cells through three-dimensional static culture and three-dimensional dynamic culture methods.The cell viability and chondrogenic ability of the two groups of microtissues were detected by scanning electron microscopy,live and dead staining,and RT-qPCR. RESULTS AND CONCLUSION:(1)Cartilage-derived microcarriers were successfully prepared.Compared with before decellularization,the DNA content significantly decreased after decellularization(P<0.001).Scanning electron microscope observation showed that the surface of the microcarrier was surrounded by collagen,maintaining the characteristics of the natural extracellular matrix of cartilage cells.CCK-8 assay indicated that microcarriers had no cytotoxicity and could promote cell proliferation.(2)Scanning electron microscopy and live and dead staining results showed that compared with the three-dimensional static group,the three-dimensional dynamic group had a more extended morphology of microtissue cells,and extensive connections between cells and cells,between cells and matrix,and between matrix.(3)The results of RT-qPCR showed that the expressions of SOX9,proteoglycan,and type Ⅱ collagen in microtissues of both groups were increased at 7 or 14 days.The relative expression levels of each gene in the three-dimensional dynamic group were significantly higher than those in the three-dimensional static group at 14 days(P<0.05).At 21 days,the three-dimensional static group had significantly higher gene expression compared with the three-diomensional dynamic group(P<0.001).(4)The results showed that compared with three-dimensional static culture microtissue,three-dimensional dynamic culture microtissue could achieve higher expression of chondrogen-related genes in a shorter time,showing better cell viability and chondrogenic ability.

This study was designed to explore the immune reconstitution efficacy of human thymic slices transplanted into renal capsule,subcutaneous or muscle of nude mice,and further explore the optimal location of heterotopic transplantation.The thymus tissue discarded from congenital heart disease patients was made into 0.5-1 mm thick tissue sections and cultured in vitro to remove immune cells.H&E staining and immunohistochemical staining were used to assess the residual tissue structure and cell types in thymic slices,while quantitative PCR methods were used to assess the function of residual cells in thymic slices.Then thymic slices were transplanted into the renal capsule,subcutaneous or muscle of nude mice,and the immune reconstitution efficacy was compared by flow cytometry and histology.Data showed that after 14 days of culture in vitro,the clearance rate of T lymphocytes in the thymic slices was more than 90%,and the epithelial cell network structure of the tissue was intact,while a large number of macrophages,dendritic cells and endothelial cells remained.Quantitative PCR results showed that the gene expression levels of epithelial cell markers and secreted cytokines in cultured thymic slices could be effectively maintained.Flow cytometry showed that at 16 weeks after transplantation,the proportion of T cells in peripheral blood of mice in different transplantation groups were significantly increased,whereas the proportion of T cells in muscle group was the highest.In situ histological examination showed that the regeneration of thymus tissue was detected at all three transplant sites.In addition,the graft detection rate was 40%in the renal capsule group,60%in the subcutaneous group and 100%in the musclegroup.In conclusion,the human thymic slices cultured in vitro for 14 days retain a complete thymic matrix microenvironment.Transplantation of human thymic slices can effectively reconstruct the ratio of T cells in nude mice,and the muscle is the most effective transplantation site.

Objective:Based on gene expression omnibus(GEO),differential expressed genes,gene set enrichment analysis(GSEA)and immune cell infiltration analysis were performed on microarray data of chronic spontaneous urticaria(CSU)expression profile,to gain more insight into the pathogenesis of CSU.Methods:The GSE72541 raw data were obtained from the GEO.Differential expressed genes were screened using R software.String database were used to construct the the protein-protein interaction(PPI)net-work.Gene ontology(GO)and Kyoto encyclopedia of gene and genomes(KEGG)enrichment analysis were performed using GSEA software.The ssGSEA method was used to analyze the infiltration of immune cells in the expression profile.Results:Genes closely related to platelet activation and its function were up-regulated in CSU serum,while genes related to Th1 cell chemotaxis were down-regulated in CSU serum.Biological processes and signal pathways related to coagulation cascade reaction,regulation of vascular per-meability,immune and inflammatory reactions,and mood-modulating were up-regulated in CSU group.Immunized cell infiltration analysis showed that activated B cells,immature B cells,follicular helper T cells,and Th2 cells were down-regulated in the CSU serum.Conclusion:Platelet activation,coagulation cascade reaction and the imbalance of Th1/Th2 immunity play important roles in the pathogenesis of CSU.

Objective:To analyze clinicopathological characteristics of trauma-related melanoma and their relationship with the prognosis of patients.Methods:Clinical data were collected from 87 cases of trauma-related melanomas in Department of Dermatology, Xijing Hospital, the Fourth Military Medical University from 2009 to 2020, and their clinicopathological characteristics were retrospectively analyzed. Mann-Whitney test was used to analyze the difference in Breslow thickness of tumors between patients of different ages or genders; Spearman rank correlation analysis was used to analyze the correlation between the time from trauma to the notice of melanoma and Breslow thickness, Kaplan-Meier survival analysis and Log-Rank test were employed to analyze the relationship between clinicopathological characteristics of melanoma and the prognosis of patients; Cox regression model was used to analyze risk factors for survival duration of patients with trauma-related melanoma.Results:Among the 87 patients with trauma-related melanoma, 47 (54.02%) were males and 40 (45.98%) were females. Among them, melanoma occurred in 50 (57.47%) cases after sharp injuries, and in 37 (42.53%) after blunt injuries. In addition, 31 (35.63%) cases presented with primary lesions on the hands, and 48 (55.17%) on the feet. The Breslow thickness of the primary tumors was significantly higher in the group aged > 55 years than in the group aged ≤ 55 years ( U= 623.500, P= 0.010) , but there was no significant difference between patients of different genders ( P= 0.138) . The time from trauma to the notice of melanoma was negatively correlated with the Breslow thickness of tumors ( r=-0.203, P= 0.037) . The age of patients, Breslow thickness of tumors, Ki67 proliferation index and genetic background of tumor significantly affect the survival duration of patients with trauma-related melanoma ( P= 0.011, 0.031, 0.002 and 0.031, respectively) ; the gender, type of trauma and ulceration of tumor mass did not significantly affect the survival duration of patients ( P= 0.618, 0.114 and 0.379, respectively) . Cox regression model analysis showed that the Ki67 proliferation index and Breslow thickness were independent risk factors affecting the prognosis of trauma-related melanoma (risk ratio [ RR] and 95% confidence interval [ CI] were 1.946 (1.234, 4.217) and 1.839 (1.014, 3.332) , P= 0.039 and 0.045, respectively) . Conclusion:The Breslow thickness of trauma-related melanoma is related to the age of patients and time from trauma to the notice of melanoma; the age, Breslow thickness of tumors, Ki67 proliferation index and genetic background of tumor all affect the survival duration of patients with melanoma, and Ki67 proliferation index and Breslow thickness are independent risk factors affecting prognosis.

Adaptation, Psychological ; Adolescent ; COVID-19 ; Child ; China ; Cross-Sectional Studies ; Female ; Humans ; Male ; SARS-CoV-2

OBJECTIVE: To report on a case with homozygous deletion of large β gene cluster and its clinical characteristics. METHODS: A total of 71 001 peripheral blood samples were subjected to capillary electrophoresis and conventional testing for common thalassemia mutations. The genotypes of suspected β gene cluster deletions were analyzed by Gap-PCR and multiplex ligation-dependent probe amplification (MLPA). Their hematological characteristics were compared by statistical analysis R software. RESULTS: Eighty-nine cases were detected with Chinese CONCLUSION The carrier rate for large fragment deletions of β gene cluster in Huizhou region is rather high, for which the value of HbF is significantly increased. Attention should be paid to screening and diagnosis of rare genotype to prevent missed diagnosis and/or misdiagnosis.
Gene Deletion ; Homozygote ; Humans ; Multigene Family/genetics* ; Phenotype ; beta-Thalassemia/genetics*

Objective To evaluate the efficacy of in-situ split liver transplantation (ISSLT) in children.Methods From June 2015 to August 2018,10 liver grafts from DBD were split in-situ.All the donors were male,and the median age of the donors was 28.5 year old (18-48 year).One left half graft and 9 left lateral lobe grafts (including 2 reduced size grafts) were transplanted to 10 pediatric recipients.Four grafts were transplanted in our center,and the rest 6 grafts were shared to other two transplant center.The primary diseases of the recipients included biliary atresia (8/10),hepatic sinus obstruction syndrome (1/10) and Alagille syndrome (1/10).The median age of the recipients was 10 month (7 month-11 year),and the mean body weight was 9.8 ± 6.6 kg (5-28 kg).Results All liver grafts were split in-situ.The mean split time of liver grafts was 88.5 ± 18.9 min.The mean weight of split grafts was 336.7-± 85.4 g.All recipients were subjected to piggyback liver transplantation.Operation time was 542.5 ± 112.1 min.Anhepatic time was 52.0 ±-13.5 min.GRWR was (3.98 ±0.96)％.GRWR of two cases was more than 5％,so segment Ⅲ was partially reduced.During the follow-up period,9 cases were alive and 1 case died due to multiple organ failure 1 day after liver transplantation.Conclusions ISSLT can enlarge the graft pool for children and achieve good results.

OBJECTIVETo explore hematological and molecular characteristics of Hemoglobin Q-Thailand in Huizhou area of Guangdong Province.

METHODSA total of 34 977 samples were screened by capillary and agarose gel electrophoresis. Samples suspected with HbQ strips were subjected to blood cell count and DNA sequencing. Twenty three common mutations associated with α- and β-thalassemia were identified by liquid phase chip and diversion hybridization technique.

RESULTSThe carrier rate of Hb Q-Thailand in Huizhou area was 0.13%. Pedigree analysis indicated that the Hb Q-Thailand allele is linked with a leftward single a-globin gene deletion (-α). Hematological index (HGB, MCV, MCH, HbA, HbA, HbQ) of 45 heterozygous carriers of Hb Q-Thailand were (130.25±17.37) g/L, (79.81±4.97) fl, (26.38±1.48) pg, (71.37±5.07)%, (1.65±0.45)%, (26.87±4.95)%, respectively. A statistical difference was also found in their hematological index of HbA and HbA compared with 408 heterozygous carriers of -α mutation (P<0.05).

CONCLUSIONHb Q-Thailand has a high detection rate in Huizhou area. The allele is mainly in a heterozygous status and linked with -α. The Hb Q strip can be detected by hemoglobin electrophoresis. When combined with other types of thalassemia, the heterozygotes will show unique hematological parameters.

Objective To observe the changes of abdominal oxygen saturation in very low birth weight infants (VLBWI)with feeding intolerance (FI)within 1 4 days after birth monitored by near infrared spectroscopy (NIRS).Methods VLBWI fitting entry criteria were enrolled into this study.NIRS monitoring was carried out to detect cerebral oxygen saturation (ScO2 )and abdominal oxygen saturation (SsO2 ).Data were analyzed between FI infants and feeding tolerance (FT)infants.FI was defined as follows:gastric residual of more than 50% of the previous feeding volume;emesis or abdominal distention or both;decrease,delay or discontinuation of enteral feedings. Results 93 VLBWI were enrolled.52 cases(55.91 %)presented with FI,including 29 cases(31 .1 9%)of gastric residual increasing and 23 cases(24.73%)of emesis with or without abdominal distention within 1 4 days after birth. The levels of SsO2 and SsO2 /ScO2 showed no differences in infants with FT and with FI within 24h after birth (P >0.05).The change rates of the median of SsO2 and SsO2 /ScO2 in FT infants were similar during 1 4 days (P >0.05).While both the change rates of SsO2 and SsO2 /ScO2 were markedly decreased 1 day before and the day of FI (P <0.01 ).The decreasing degree of SsO2 was similar between infants with gastric residual increasing and infants with emesis with or without abdominal distention[(1 6.2 ±5.1 )vs (1 7.4 ±3.6)%,t =0.733,P =0.476]. Conclusion Abdominal oxygen saturation measured by NIRS may be a useful method for infants adjusting the feeding plan.

Objective To analyze the correlation between serum hs -CRP levels and MR Imaging in ischemic cerebrovascu-lar patients.To explore the hs -CRP serum levels in ischemic cerebrovascular patients incidence of early diagnosis and prognosis e-valuation application value.Methods 300 cases of ischemic cerebrovascular disease as the research object were selected from May 2013 to December 2014.The disease is classified into acute cerebral infarction group 100 cases, acute phase of cerebral infarction group 140 cases and transient ischemic attack 60 cases.Among 100 cases of patients with acute cerebral infarction group, in accord-ance with the MR Imaging data, they can be subdivided into three groups.Group A: lacunar infarction group 28 cases; Group B:small area of infarction group 50 cases and group C: large area of infarction group 22 cases.Another 40 healthy physical examination were selected as control group.The hs -CRP serum levels were made correlation analysis with MR Imaging.Results The acute cerebral infarction and transient ischemic attack group serum hs -CRP level difference compared with the control group with statisti-cal significance (p<0.05); The SPSS Spearman correlation analysis result shows that the hs -CRP serum levels and MR Imaging data shown in the infarction area and the body has a direct correlation .Conclusion The ischemic cerebrovascular disease patients se-rum hs-CRP level with MR Imaging have shown increased infarct size and raise.Damage area is bigger, and the corresponding serum hs-CRP levels changes more violently.