Antiphospholipid Syndrome ; pathology ; physiopathology ; Child ; Female ; Follow-Up Studies ; Humans

BACKGROUND:Conventional methods,including trypsin digestion and cells transfer using single call suspension,have many drawbacks,which limit the development of bone tissue engineering.OBJECTIVE:To culture bone marrow stromal stem calls,induce osteoblastic differentiation,and prepare cell sheets.METHODS:Canine bone marrow stromal calls were isolated by density gradient centrifugation technique,inoculated into DMEM medium,and induced to differentiate into osteoblasts.Complete call sheets were harvested by call sheet engineering based on the temperature change of temperature-responsive medium.RESULTS AND CONCLUSION:Immediately after inoculation,primary calls were scattered on the bottom of culture flask,presenting a transparent spherical body with a good refractive capacity.At 12 hours,calls exhibited a long shuttle shape,reached complete confluency,and grew in a whirlpool-like fashion.After osteoblastic induction,the majority of bone marrow stromal stem calls appeared tetragonal,polygonal,and squamose.At 21-28 days,round or oval-shaped calcified nodules formed.When the bone marrow stromal stem calls in the temperature-responsive culture dishes were cooled below the critical temperature 32℃,cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem call sheets.These findings indicate that density gradient cantrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and call sheet engineering enables to harvest complete bone marrow stromal stem call sheets.

BACKGROUND:There are some disadvantages in harvesting and transferring cells in the traditional tissue engineering technique,and it is difficult to form dense tissues,which significantly limits the development of tissue engineering.OBJECTIVE:To explore the culture and fabrication of dog bone marrow mesenchymal stem cell(BMSC)sheet in vitro.METHODS:Bone marrow was extracted from dogs following anesthesia.BMSCs were isolated with the method of density gradient centrifugation in vitro.BMSCs at passage 4 at a density of 1×10~9/L were incubated in the temperature-responsive culture dishes with a diameter of 3.5 cm,and cultured in an incubator at 37 ℃,5% CO_2 and saturated humidity.The temperature of the incubator was changed from to 37 V to 20 ℃ to prepare BMSCs cell sheet for 20 minutes.Cell morphological changes and cell sheet formation were observed under an inverted microscope.RESULTS AND CONCLUSION:Dog BMSCs following 24 hours of primary culture presented ellipse or polygonal shape.Most cells adhered at hour 72,and cell colonies were visible at day 7.Cells showed long spindle and completely confluence at day 12,with unclear boundary.BMSCs in the temperature-responsive culture dishes presented short spindle shape,and gradually separated from the dish bottom,forming entire cell sheet containing extracellular matrix at 20 V.These verified that dog BMSCs can be effectively obtained with method of density gradient centrifugation.Complete cell sheet layer can be fabricated with temperature-responsive culture dishes.

BACKGROUND: How to reconstruct tissue-engineered bone with structure similar to natural bone iS a problem in the development of tissue engineering. Cell sheet engineering technology enables novel approaches to construction of tissue-engineered bone. OBJECTIVE: To observe the biocompatibility of call sheets to decalcified bone matrix (DBM) and their growth on DBM. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory, Affiliated Hospital, Qingdao University Medical College between June and September 2009.MATERIALS: Dog bone marrow stromal cell sheets were prepared using temperatura-responsive medium. Dog DBM was prepared by defatting, decalcification, and noncotlagen protein removal procedures. METHODS: DBM surface was covered by call sheets prepared by temperature-responsive technology and cultured with DMEM containing 10% fetal bovine serum and osteoinductive agent.MAIN OUTCOME MEASURES: Under scanning electron microscope, DBM structure, as well as the attachment and growth of cell sheets on DBM surface, was observed. Porosity and aperture size of DBM were calculated. RESULTS: DBM exhibited a three-dimensional latticed structure, with a porosity of approximately 75%. The mean aperture size was (250.11±98.89) μm, exhibiting a normal distribution. Cell sheets well attached to and grew on DBM surface, and rapidly proliferated.CONCLUSION: Cell sheets show good biocompatibility to DBM. DBM/cell sheets complex can be applied in tissue-engineered bones, which promotes the construction of tissue-engineered bone with structure similar to natural bone.

Cytomegalovirus (CMV) retinitis (CMVR) is a common opportunistic infection of the eye after allogeneic hematopoietic stem cell transplantation in patients with hematological diseases. It often occurs within 3 months after the operation, with CMV activation and high blood CMV peaks. It often occurs on patients with long-term CMV viremia, human leukocyte antigen incompatible transplantation, unrelated donor transplantation, haploid transplantation, childhood hematopoietic stem cell transplantation, delayed lymphocyte engraftment, acute and chronic graft-versus-host disease after surgery. The visual prognosis of patients is related to the area of CMVR lesions on the retina, the number of quadrants involved, whether the macula is involved, and the CMV load of the vitreous body is involved, and it is not related to whether the Epstein-Barr virus infection is combined with blood and vitreous humor. The incidence of CMVR is increasing year by year. It is helpful that paying attention to systemic risk factors and epidemiology can provide more effective guidance for ophthalmologists during diagnosis and treatment, help patients improve the prognosis of vision, and reduce or even avoid the occurrence of blindness caused by CMVR.

Objective To make comparisons of the three models of acute and chronic rheumatic carditis to find out an optimal animal model.Methods AntigenⅠwas a emulsifier mixed by complete freund’ s adjuvant( CFA) and Group A streptococcus(GAS).AntigenⅡwas mixed by incomplete freund’s adjuvant(IFA) and GAS.Female Lewis rats were randomly divided into four groups: A, B, C treatmeat groups were immuned with antigenⅠat the foot pad firstly. Subsequently, rats in group A、B、C were injected antigenⅠ, antigenⅡand activated GAS respectively to make the models of RHD.Rats in control group D were immunized with the same protocol outlined as treatment groups but without GAS. Respectively 7, 12, 24 weeks the rats were sacrificed 24 ( each group was 6).The blood biochemical item and Hematoxylin-eosin( HE) staining of hearts were detected.Results In group C the mortality was 25%.In group A, the incidence of carditis was the highest.Histopathological manifestations of group A, C was not only revealed acute damage such as inflammatory cell infiltrate as well as group B, but also the Aschofflike cells in the myocardial cells interstitial.But in group A and C there had a great degree of the inflammatory cells infiltration than group B.At 24th week rats in group A detected the rate and degree of valve fibrosis in chronic damage were higher than group B and C.None of rats in group D presented carditis or valvulitis.Conclusion In group A, giving the GAS with continuous stimulation after using the mixed emulsification of CFA and GAS to immune Lewis rats for five times was a appropriate method which could provide an optimal animal model for experimental study of acute and chronic rheumatic heart disease.

Objective To discuss the possible molecular mechanisms involved in the protective effects of Biifdobacterium on intestinal tissue of necrotizing enterocolitis (NEC) newborn rats. Methods Seventy-five newborn Sprague-Dawley rats (born within 2 h) were randomly divided into five groups, each group with 15 rats. Group A was the NEC model group, and the rats were fed lipopolysaccharide (LPS) and formula. Group B was the Biifdobacterium treatment group, and the rats were fed LPS and formula and Biifdobacterium micro-capsule. Group C was the artificial feeding control group, and the rats were fed formula. Group D was the Biifdobacterium control group, and the rats were fed formula and Biifdobacterium micro-capsule. Group E was the breastfeeding control group, and the rats were fed rat breast milk by mothers. LPS 30 mg/kg was administered by gavage once per day for 3 days. Bifidobacterium micro-capsules were given as 1×1010 colony forming units/ml by gavage with formula once per day. After fed for 72 h and fasted for 12 h, the five groups of rats were killed by decapitation. Morphological changes in the terminal ileum tissue were observed under a light microscope and intestinal injury was scored. The expression of Toll-like receptor (TLR) 2, TLR4, and nuclear transcription factor (NF)-κB p65 was detected by immunohistochemical methods. Kruskal-Wallis test, analysis of variance, corrected Chi-square test and Fisher's exact test were used for statistics. Results The morbidity of NEC in group A to E was 11/15, 4/15, 3/15, 2/15 and 0/15, respectively;the intestinal injury score in group A to E was 3.37±0.27, 1.53±0.44, 1.75±0.37, 0.92±0.39 and 0.30±0.18, respectively; the expression level of TLR2 in group A to E was 0.35±0.05, 0.30±0.03, 0.32±0.04, 0.30±0.02 and 0.29±0.03, respectively;the expression level of TLR4 in group A to E was 0.48±0.05, 0.34±0.03, 0.36±0.03, 0.37±0.04 and 0.35±0.02, respectively;the expression level of NF-κB p65 in group A to E was 0.43±0.03, 0.29±0.03, 0.35±0.02, 0.32±0.02 and 0.30±0.02, respectively. The differences in NEC morbidity, intestinal injury score, and the expression levels of TLR4, TLR2 and NF-κB p65 among the five groups were all statistically significant (χ2, H or F=23.863, 70.290, 8.803, 38.599 and 75.076, respectively, all P<0.05). The values in the NEC model group were all significantly higher than those in the other four groups (all P<0.05). The morbidity of NEC in the Biifdobacterium treatment group compared with the three control groups was not significantly different (all P > 0.05). The intestinal injury score in the Bifidobacterium treatment group was significantly higher than that in the Bifidobacterium control group and the breastfeeding control group (both P < 0.01), but was not significantly different to that in the artificial feeding control group (P > 0.05). The expression levels of TLR4 and NF-κB p65 in the Biifdobacterium treatment group were significantly lower than those in the artificial feeding control group and the Biifdobacterium control group (all P < 0.05), and were not significantly different to those in the breastfeeding control group (P>0.05). The expression level of TLR2 in the Biifdobacterium treatment group compared with the three control groups was not significantly different (all P > 0.05). Conclusions Biifdobacterium may inhibit pathogenic bacteria or regulate the negative feedback of TLR2 to reduce the expression of TLR2 and TLR4 in intestinal mucosa cells, inhibit the NF-κB pathway, attenuate the inflammatory reaction, and play a role in the prevention and control of NEC.

Objective To investigate the effects of circumferential pulmonary vein isolation (CPVI) on atrial effective refractory period (ERP) in patients with paroxysmal atrial fibrillation.Methods 30 patients who underwent radiofrequency catheter ablation for paroxysmal AF were enrolled in this study.Using FAM mode,the RA and LA anatomical models were achieved in the CARTO 3 system.SVC,MRA,RAA,LA-A,LA-R,LA-P,LAA,LSPV,LIPV,RSPV,RIPV,CSp,CSd,were respectively located in the RA or LA anatomical model.Before and after CPVI,ERPs were measured in different locations of the atrium using programmed stimulation.The ERPs of the RA (SVC,MRA,RAA,CSp),LA (LA-A,LA-R,LA-P,LAA,CSd),PVs (LSPV,RSPV,LIPV,RIPV) were compared.Bilateral CPVIs were completed in all patients,and PV-LA bidirectional conduction block was achieved.The changes of electrophysiological characteristics of atrium before and after CPVI were observed.Results (1) ERP at different locations in the atrium before CPVI:Comparisons of ERPs at different locations of atrium:RAA had the minimal ERPs[(197.4 ± 28.6) ms (P ＜ 0.01);followed by PVs measuring,respectively,LSPV (213.0 ± 47.5) ms,LIPV (208.9 ± 45.9) ms,RSPV (209.3 ± 43.6) ms,RIPV (213.5 ± 48.1) ms and LAA (218.1 ± 27.7) ms.Comparisons of ERPs in RA,LA,and PVs showed:PVs had the lowest ERPs (211.2 ± 35.2) ms versus RA ERP (227.0 ± 23.7) ms versus LA ERP (241.0 ± 21.5) ms (P ＜ 0.05).(2) Comparisons of ERPs before and after CPVI:Comparisons of ERPs at different locations of atrium showed:RAA [(197.4 ± 28.6) ms vs.(208.6 ± 32.2) ms,P=0.003],CSp [(234.7 ± 29.1) ms vs.(246.9 ± 29.7) ms,P=0.007],LA-R [(242.9 ± 28.9) ms vs.(258.3 ± 26.9) ms,P=0.003],LA-P [(252.2 ± 28.5) ms vs.(261.1 ± 30.2) ms,P=0.039]and CSd [(238.6 ± 28.3) ms vs.(250.3 ± 23.6) ms,P =0.009].ERPs were found statistically prolonged at all different locations after CPVI.Comparisons of ERPs at RAand LA after CPVI showed:RA [(227.0 ± 23.7) ms vs.(235.9 ± 21.7)ms,P=0.002]and LA [(241.0 ± 21.5) ms vs.(249.7 ± 19.9) ms,P =0.001],which were statistically increased after CPVI.(3) A total of 90 episodes of atrial arrhythmias were induced before CPVI which were found at RAA (n =17),LAA (n =12),and PVs (n =36).After CPVI,8 episodes of atrial arrhythmias were induced which were found at,RAA (n =4),LAA (n =3),and SVC (n =1).Conclusions (1) Compared with other parts of atrium,ERPs at PVs,LAA and RAA are significantly shorter in patients with paroxysmal AF.At PVs,LAA and RAA,atrial arrhythmias are easily to be induce by programmed stimulation.(2) In patients with paroxysmal Af:PVs has the shortest ERPsfollowed by RAs whereas LA ERPs is the longest.There is a large ERP gradient change between PVs and LA.(3) The ERPs at RAs,LAs,As,and LA-PV are prolonged after CPVI.(4) Atrial arrhythmia is less likely to be induced after CPVI.

Objective:To summarize the clinical experiences of pediatric en-bloc kidney transplantation (EBKT) at a single center and explore the measures of improving its efficacy.Methods:Clinical data and outcomes retrospectively analyzed for 38 EBKT children between September 2014 and November 2019 from Department of Urology Affiliated Union Hospital Tongji Medical College Huazhong University of Science & Technology. The pediatric donors were aged (63.6±5.7) days with a weight of (4.1±0.2) kg. And the transplant recipients were aged (28.1±1.4) years with a weight of (48.7±4.9) kg. Serum levels of creatinine and basic profiles of both donors and recipients were recorded at Day 0/7/30/90/80/360 post-EBKT. The postoperative occurrences of such complications such as thrombosis, urine leakage, delayed graft function (DGF), proteinuria and hematoma were measured.Results:The one-year graft survival rate was 76.3%(29/38) and the recipient survival rate 100.0%(38/38). Among 29 recipients with long-term graft survival, no dialysis was required at Week 2 post-EBKT and the serum level of creatinine dropped to normal at Year 1. Thrombosis was a major post-EBKT complication with an incidence of 18.4%(7/38). The other complications included urine leakage (20.7%, 6/29), hematoma (6.9%, 2/29) and primary non-functioning kidney (2.6%, 1/38).Conclusions:As an effective way of expanding the pool of donors, pediatric EBKT is clinically feasible.

Objective : To investigate the awareness of nutritional health knowledge regular unpaid blood donors in Zhoushan City, Zhejiang Province, so as to provide insights into nutrition and health education among unpaid blood donors. Methods: The regular unpaid blood donors were sampled using a convenient sampling method from Zhoushan Central Blood Station from January 1, 2021 to June 30, 2022. The nutritional health knowledge was investigated using the Chinese Nutrition Health Knowledge Questionnaire for Adults compiled by Chinese Center for Disease Control and Prevention, and the awareness and source of nutritional health knowledge was descriptively analyzed. Results: Totally 526 questionnaires were allocated, and 502 valid questionnaires were recovered, with an effective recovery rate of 95.44%. The respondents included 240 men (47.81%) and 262 women (52.19%), and included 343 individuals at ages of 18 to 44 years (68.33%), and 159 individuals at ages of 45 years and older (31.67%). The overall awareness of nutritional health knowledge was 14.54% among regular unpaid blood donors in Zhoushan City, and a higher awareness was seen among female regular unpaid blood donors (17.56%) than among males (11.25%) (P<0.05), while the awareness of nutritional health knowledge was significantly higher among respondents at ages of 18 to 44 years than among those at ages of 45 years and older (16.91% vs. 9.43%, P<0.05). The awareness of nutritional health knowledge was significantly higher among respondents with an educational level of junior college and above than among those with an educational level of high school/technical secondary school/technical school (17.24% vs. 12.22%, P<0.05), and the awareness was significantly higher among respondents with healthcare-related occupations than among those with other occupations (16.44% vs. 14.22%, P<0.05). Wechat, Weibo, Tik Tok and Kuaishou were main routes to obtain nutritional health knowledge (83.86%). Conclusions The awareness of nutritional health knowledge is low among regular unpaid blood donors in Zhoushan City. Men, middle-aged and elderly residents and residents with a low educational level are targets that should be given a high priority for nutritional health education, and new media may be fully utilized for nutritional health education.