Objective To study the feasibility of TNF-α promoting migration of rat mesenchymal stem cells (MSCs) to local damaged tissues. Methods The MSCs was exposed to TNF-α at different concentrations and the expression rate of surface adheslon molecules and specific markers as well as their adhesion to endothelial cells were detected.Based on the above steps,the MSCs stimulated with the optimal concentration of TNF-α were obtained and were injected intravenously to the rats whose hindlimbs experienced ischemia damage.The rats were executed for achieving the muscle samples in the ischemic area,which were made into frozen section to count the number of MSCs. Results ( 1 ) Twenty-four hours after the TNF-o stimulation,the expression of adhesion molecule (VCAM-1) of MSCs increased in a concentration-dependent manner,while the expression of adhesion molecules (ICAM-1,L-Selectin and VLA4) of MSCs showed no significant changes.Besides,the expression rate of specific markers of MSCs was also obscure.(2) Exposed to 10 ng/ml TNF-o,MSCs presented an obviously increased ability in adhesion to the endothelial cells.(3) MSCs stimulated with 10 ng/ml TNF-α showed a larger number in the ischemia-damaged tissue of rat hindlimbs than that in the control group. Conclusion TNF-α at concentration of 10 ng/ml is effective within a short term in increasing VCAM-1 expression in rat MSCs and promoting the adhesion of MSCs to endothelial cells without affecting their character.

BACKGROUND: Following bone marrow mesenchymal stem cells (BMMSCs) infusion therapy, which factor promotes BMMSCs migrated to correct position is a key point, currently, adhesion molecule is thought to be playing an important role in mediating BMMSCs migration. OBJECTIVE: To investigate the expression of vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) in rat BMMSCs. METHODS: BMMSCs were in vitro separated from rat bone marrow by directly adherence method. The expression of VCAM-1 and ICAM-1 were identified by using immunocytochemical staining, and the expression rates of antigen were tested by flow cytometry, in addition, their mRNA expressions were measured by RT-PCR. RESULTS AND CONCLUSION: Immunocytochemistry demonstrated that BMMSCs weakly expressed VCAM-1, but strong expressed ICAM-1. Flow cytometry showed that the expression rate of VCAM-1 was 6%, and the expression rate of ICAM-1 was 100%. RT-PCR showed that BMMSCs expressed a low level of VCAM-1 mRNA but a high level of ICAM-1 mRNA. It revealed under physiological condition, BMMSCs expressed a low level of VCAM-1, whereas they expressed a high level of ICAM-1.

Objective By studying the variation of individual urinary iodine concentration due to different ways of urine sample collection to optimize it for standard clinical evaluation.Methods Totally 20 healthy adults were recruited and their urine samples were collected as a random urine sample within 1 day,the 24 hour urine and morning urine samples within 5 successive days,respectively.The coefficient of variation in each group was calculated.Paired t test was used to compare the results of 24 hour urine with the results of random urine and that of morning urine samples,respectively.Results The range of individual coefficient of variation for random urine sample within one day was 12.5％-57.4％,while most of the coefficients of variation were around 30.39％.In contrast,the individual coefficients of variation of morning urine sample and 24 hour urine results within 5 days were 5.4％-26.0％ and 3.4％-16.6％ and most of them were at about 11.74％ and 7.91％.The paired t test showed that the results of random urine sample were significantly different compared with that of 24 hour urine (t =-4.231,P ＜ 0.05).On the other hand,there was also significant difference for the results of morning urine compared with that of 24 hour urine (t =3.884,P ＜ 0.05).Conclusion This study suggests that 24 hour urine is the most appropriate way of sample collection for individualized detection of urinary iodine.

Objective To investigate whether the percutaneous ethanol injection (PEI)under sedation and analgesia can in-crease the energy deposition and curative efficiency of the high intensity focused ultrsound(HIFU)in treating unresectable middle and advanced stages of primary liver cancer.Methods Thirty-six cases of clinically diagnosed unresectable middle and advanced sta-ges of primary liver cancer were randomly divided into the PEI+ HIFU group(combination group,n = 23)and the simple HIFU group (HIFU group,n=13);10mL of the mixture of 99.7% ethanol and iodized oil (9:1)was given by intratumoral injection at 30 min before ablation in the PEI+HIFU group,while 0.9% physiological saline 10mL was replaced in the simple HIFU group.The ablation energy efficiency factor(EEF)and irradiation time were compared between the two groups.Results The ablation EEF in the PEI+HIFU group and the simple HIFU group were (13.82+4.26)J/mm3 and (25.63+6.31)J/mm3 respectively,the PEI+HIFU group was significantly lower than the simple HIFU group (P <0.05);the irradiation time were (1 468.28+253.21)s and (2 352.56+463.34)s respectively;which in the PEI+ HIFU group was significantly shortened (P <0.05).Conclusion PEI can enhance the HIFU ablation energy deposition and improve the efficiency of HIFU for treating unresectable primary liver cancer.

BACKGROUND:Although the clinical application of Masquelet technology has achieved extensive success,the research on optimizing all aspects of Masquelet technology is still being carried out.The focus of doctors is to speed up bone healing and shorten bone healing time after bone grafting. OBJECTIVE:To observe the effect of calcium phosphate combined with recombinant human bone morphogenetic protein-2 in repairing tibial infectious bone defects. METHODS:Thirty-one patients with tibial infectious bone defects were selected from The People's Hospital of Jianyang City from June 2017 to June 2022.They were treated with the Masquelet membrane induction technique.During the second stage of operation,they were divided into a control group(n=15)and a study group(n=16)according to different bone graft materials.Patients in the control group were implanted with autologous bone/allogeneic bone particles,and those in the study group were implanted with calcium phosphate combined with recombinant human bone morphogenetic protein-2/autologous bone particles.Six months after the second stage operation,peripheral blood inflammatory indexes such as white blood cell count,C-reactive protein,and erythrocyte sedimentation rate were detected.Imaging bone healing time,bone healing X-ray score,bone defect healing classification,and adjacent joint function were recorded.The presence of nail track infection,implant absorption,pain,and infection in the bone extraction area were observed. RESULTS AND CONCLUSION:(1)White blood cell count,erythrocyte sedimentation rate,and C-reactive protein levels of the two groups 6 months after the second stage operation were significantly lower than those before the first stage operation(P<0.05).There was no significant difference in each index between the two groups(P>0.05).(2)Bone healing time in the study group was shorter than that in the control group(P<0.05).(3)The Samantha X-ray score of the study group 6 months after the second stage operation was higher than that of the control group(P<0.05).The excellent and good rate of bone defect healing and adjacent joint function of the study group was higher than that of the control group(P<0.05).There was no significant difference in the recurrence rate and complication rate between the two groups(P>0.05).(4)These findings indicate that the effect of calcium phosphate combined with recombinant human bone morphogenetic protein-2 during the second stage operation of the Masquelet membrane induction technique in the treatment of tibial infectious bone defect is good and safe.

BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.