A retrospective study of correlation between serum levels of omega-3 polyunsaturated fatty acids and post-stroke cognitive impairment / 中华物理医学与康复杂志

Objective:To quantify any correlation between serum levels of omega-3 polyunsaturated fatty acids (ω3-PUFAs) and post-stroke cognitive impairment (PSCI).Methods:The clinical data of 77 patients hospitalized after a first stroke were analyzed. The Minimum Mental State Examination (MMSE) was used to divide them into impaired (PSCI) and unimpaired (non-PSCI) cohorts. The serum levels of ω3-PUFAs, α-linolenic acid (ALA), eieosapentaenoic acid (EPA) and dueosahexenoie acid (DHA) were compared between the two groups and correlated with the individuals′ MMSE scores.Results:The average ALA, EPA, DHA and total ω3-PUFAs levels of PSCI group were in most cases significantly lower than those of the non-PSCI group. Spearman correlation analysis showed that serum DHA level was a weak positive predictor of the MMSE scores (R=0.32, P≤0.05). Logistic regression analysis indicated that low serum DHA level was an independent risk factor for PSCI ( P≤0.01). Conclusions:Cognitively impaired stroke survivors tend to have lower serum ω3-PUFAs levels than those without cognitive impairment. There is a weak positive correlation between serum DHA levels and MMSE scores. Low serum DHA level is an independent risk factor for PSCI. The serum level of ω3-PUFAs is of high value in the auxiliary diagnosis and evaluation of PSCI.

A multicenter clinical study of 280 cases of staphylococcal peritoneal dialysis-associated peritonitis / 中华肾脏病杂志

Objective:To investigate the incidence, drug sensitivity and drug resistance characteristies, and theraputic effect of staphylococcal peritoneal dialysis-associated peritonitis (PDAP), aim to provide clinical evidences for standardizing treatment therapy of staphylococcal PDAP. Methods:Clinical data of PDAP patients admitted to the Second Hospital of Jilin University, the First Hospital of Jilin University-the Eastern Division, Jilin Central Hospital and Jilin First Automobile Work General Hospital during January 1, 2013 and December 31, 2019 were retrospectively collected. The results of etiology, drug sensitivity and drug resistance of staphylococcal PDAP patients were collected. According to the pathogenic bacteria, patients were divided into staphylococcus aureus group ( n=48) and coagulase-negative staphylococcus group ( n=232). According to the results of methicillin resistance, patients were divided into drug-resistant group ( n=71) and drug-sensitive group ( n=30). The prognosis of antibiotic therapy in each group were compared. Poisson regression was used to test the changing trend of the incidence of staphylococcal PDAP. The changes of drug sensitivity and drug resistance of staphylococcus were compared between 2013 and 2019 by linear trend χ2 test. Results:A total of 1 085 cases of PDAP occurred in 625 patients were screened, and 280 cases of staphylococcal PDAP were finally included. The incidences of staphylococcal PDAP, staphylococcus aureus PDAP and coagulase-negative staphylococcal PDAP were 0.063 times per patient year, 0.010 times per patient year and 0.053 times per patient year respectively. In addition, the incidence of PDAP caused by staphylococcus, staphylococcus aureus and coagulase-negative staphylococcus decreased year by year (all P<0.05). With the change of years, the sensitivity rate of staphylococcus to rifampicin increased, while the sensitivity rate of staphylococcus to moxifloxacin decreased (both P<0.05). The drug resistance rate of staphylococcus to levofloxacin increased ( P<0.05). The staphylococcus aureus group was more prone to refractory PDAP and catheter removal than that in coagulase-negative staphylococcus group, and the recurrence rate was higher than that in coagulase-negative staphylococcus group (all P<0.05). The proportion of vancomycin used during the whole course of antibiotic therapy in drug-resistant group was higher than that in drug-sensitive group ( P<0.05). Conclusions:The incidence of staphylococcal PDAP decreases year by year, and the drug sensitivity characteristics of staphylococcus also change. The therapeutic outcomes of staphylococcus aureus PDAP are worse than that of coagulase-negative staphylococcus.

IL6 Receptor Facilitates Adipogenesis Differentiation of Human Mesenchymal Stem Cells through Activating P38 Pathway

Background and Objectives@#Mesenchymal stem cells (MSCs) have the multipotent capacity to differentiate into multiple tissue lineages as well as to self-renew, which is the main origin of adipocytes. IL6/IL6R pathway exerts a significant role in tissue regeneration and cell differentiation. Whereas, the underlying mechanism between IL6/IL6R pathway and MSCs adipogenesis differentiation remains elusive. @*Methods@#MSCs from healthy donors were cultured in adipogenesis differentiation medium for 0∼14 days, during which their adipogenesis differentiation degree was evaluated by Oil Red O staining. The expression of IL6R was detected in MSCs during adipogenesis differentiation. Knockdown and overexpression of IL6R were respectively performed using siRNA and lentivirus to investigate its effect on MSCs adipogenesis differentiation. The adipogenesis marker genes expression and MAPK pathway activation were detected by Western blotting. The role of P38 pathway in the adipogenesis differentiation of MSCs was determined using the specific inhibitor SB203580. @*Results@#The expression of IL6 and IL6R increased during adipogenesis differentiation in MSCs, which were positively correlated with Oil Red O quantification result. Knockdown and overexpression experiments demonstrated a positive correlation between the expressions of IL6R and MSCs adipogenesis differentiation, accompanied by same trend of P38 phosphorylation. Besides, the specific P38 inhibitor SB203580 markedly inhibited the adipogenesis differentiation potential of MSCs. @*Conclusions@#This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.

Elevated TRAF4 expression impaired LPS-induced autophagy in mesenchymal stem cells from ankylosing spondylitis patients

Ankylosing spondylitis (AS) is a type of autoimmune disease that predominantly affects the spine and sacroiliac joints. However, the pathogenesis of AS remains unclear. Some evidence indicates that infection with bacteria, especially Gram-negative bacteria, may have an important role in the onset and progression of AS. Recently, many studies have demonstrated that mesenchymal stem cells (MSCs) dysfunction may contribute to the pathogenesis of many rheumatic diseases. We previously demonstrated that MSCs from AS patients exhibited markedly enhanced osteogenic differentiation capacity in vitro under non-inflammatory conditions. However, the properties of MSCs from AS patients in an inflammatory environment have never been explored. Lipopolysaccharide (LPS), a proinflammatory substance derived from the outer membrane of Gram-negative bacteria, can alter the status and function of MSCs. However, whether MSCs from AS patients exhibit abnormal responses to LPS stimulation has not been reported. Autophagy is a lysosome-mediated catabolic process that participates in many physiological and pathological processes. The link between autophagy and AS remains largely unknown. The level of autophagy in ASMSCs after LPS stimulation remains to be addressed. In this study, we demonstrated that although the basal level of autophagy did not differ between MSCs from healthy donors (HDMSCs) and ASMSCs, LPS-induced autophagy was weaker in ASMSCs than in HDMSCs. Specifically, increased TRAF4 expression in ASMSCs impaired LPS-induced autophagy, potentially by inhibiting the phosphorylation of Beclin-1. These data may provide further insight into ASMSC dysfunction and the precise mechanism underlying the pathogenesis of AS.

Application of dilution regression method in the coagulation detection of fat blood samples / 国际检验医学杂志

Objective To explore the dilution regression method of coagulation detection(PT ,APTT) in fat blood samples . Methods We collected 40 normal blood coagulation specimens (no fat blood ,no jaundice ,no hemolysis) in Yan′an hospital of Kun-ming ,then we detected the PT and APTT of the original plasma and 3-fold diluted plasma and 5-fold diluted plasma ,the we used the data both of before dilution and diluted to do the linear regression analysis ,and finally we got the regression equations of each index .we also collected 33 fat blood samples in Yanan hospital of Kunming ,which be divide into three groups through the severity of triglycerides :mild fat blood group(1 .7 mmol/L≤TG0 .05) .Conclusion Dilution regression method can be used to detect the fat blood samples in the clinical coagulation detection .

Bone marrow mesenchymal stem cells derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages / 中国组织工程研究

BACKGROUND:Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is stil unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE: To explore the immunoregulation capability of bone marrow mesenchymal stem cels from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cels from healthy donors on ankylosing spondylitis. METHODS: Bone marrow mesenchymal stem cels were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cels were used in subsequent experiments. A human monocytic cel line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cels and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6 (TSG-6) proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the mRNA level of cytokines secreted by bone marrow mesenchymal stem cels and macrophages. RESULTS AND CONCLUSION:The typical mesenchymal stem cel surface markers were expressed in both bone marrow mesenchymal stem cels from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and mRNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cels from patients with ankylosing spondylitis were obviously higher than those from healthy donors (P < 0.05). TSG-6 secreted by bone marrow mesenchymal stem cels from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cels from healthy donors in both RNA transcriptional and protein levels (P < 0.05). Our study demonstrates that bone marrow mesenchymal stem cels from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting the tumor necrosis factor-α secretion from macrophages, which reveals a mechanism of immune disorder in ankylosing spondylitis. The therapeutic mechanism of bone marrow mesenchymal stem cels from healthy donors may work by secreting enough TSG-6 to inhibit the activation of macrophages in patients with ankylosing spondylitis, and thereby to decrease the secretion of tumor necrosis factor-α. Cite this article:Sun SH, Wang P, Su CY, Xie ZY, Li YX, Li D, Wang S, Su HJ, Wu XH, Deng W, Wu YF, Shen HY. Bone marrow mesenchymal stem cels derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):13-19.

The relationship between hope level and quality of life of psoriasis vulgaris patients / 中华行为医学与脑科学杂志

Objective To explore the relationship between hope level and quality of life with psoria-sis vulgaris patients, and to make a reference for improving quality life of with psoriasis vulgaris patients. Methods 126 psoriasis vulgaris patients were investigated using the questionnaires of Herth Hope Inedx ( HHI) and Dermatology Life Quality Index( DLQI) .Results The average scores of HHI and DLQI of pso-riasis vulgaris patients were(33.37 ±5.15),(11.18 ±4.83)respectively,and the HHI score of psoriasis vul-garis patients was negatively correlated with the DLQI score ( r=-0.433, P<0.01) .The stepwise multivari-able regression showed that interconnectedness( R 2=0.177),education level( R 2=0.054) and years of psoriasis vulgaris( R 2=0.037) were the main influential factors on life quality of psoriasis vulgaris patients. Conclusion The hope level of psoriasis vulgaris patients is positively related with quality of life,so effective and targeted measures should be taken to improve their hope level in order to improve quality of life of psoria-sis vulgaris patients.

Allogeneic blood transfusion alleviates hip joint pain induced by ankylosing spondylitis / 中国组织工程研究

BACKGROUND:Pain is the main clinical manifestation for ankylosing spondylitis. At present, nonsteroid anti-inflammatory drugs are oral y taken, but the effects are limited and toxic and side effects are more. Thus, there is no effective scheme for treatment of pain induced by ankylosing spondylitis. OBJECTIVE:To investigate the correlation between postoperative joint pain al eviation and al ogeneic blood transfusion, and the mechanisms. METHODS:We retrospectively analyzed clinical data of 88 ankylosing spondylitis patients combined with kyphosis who received only one section of osteotomy surgery merging hip joint pain. We compared the visual analog scale score of hip joint and detected the variation of leucocytes, lymphocytes and immunoglobulin concentrations before and after the operation in the groups of fresh al ogeneic whole blood transfusion, autologous whole blood transfusion, and mixed transfusion of al ogeneic and autologous whole blood. Flow cytometry was used to analyze the number and ratio of peripheral blood Th17 cells and Treg cells which were both highly associated with autoimmune diseases. RESULTS AND CONCLUSION:The symptom of hip arthralgia obviously improved in both groups transfused by fresh al ogeneic whole blood or al ogeneic-autologous mixed whole blood. However, no obvious variation was detected in leucocytes, lymphocytes and immunoglobin concentration. However, flow cytometry results indicated that Th17/Treg proportion associated with autoimmune diseases was increased remarkably in peripheral blood of ankylosing spondylitis patients. Results suggested that al ogeneic whole blood transfusion can al eviate patients’ joint pain by correcting the imbalance of Th17/Treg which may improve their immune state.

The impact of burden, positive feelings, coping style and social support on caregivers' fatigue of dementia patients / 中华行为医学与脑科学杂志

Objective To explore the impact of burden,positive feelings,coping style and social support on caregivers' fatigue of dementia patients.Methods 190 caregivers of dementia patients were investigated by using the questionnaires of Caregivers Burden Inventory(CBI),Positive Aspects of Caregiving(PAC),Simplified Coping Style Questionnaire (SCSQ),Social Support Rating Scale (SSRS) and Fatigue Scale-14 (FS-14).Results The caregivers' fatigue of dementia patients was positively correlated with burden(r=0.56,P＜0.01),negative coping style(r=0.31,P ＜0.01),and negatively associated with positive feelings(r=-0.33,P ＜0.01),social support (r =-0.36,P ＜ 0.01) and positive coping style (r---0.55,P ＜ 0.01),respectively.Path analysis showed coping style and social support had direct impacts on fatigue; burden had a direct impact on fatigue,and also had an indirect influence on it through coping style or social support; fatigue might be decreased by positive feelings and increased by negative coping style.Conclusion To prevent and relieve the caregivers'fatigue of dementia patients,caregivers should be encouraged to take positive coping style and increase social support so as to reduce burden and increase positive feelings.

Characteristics of depression in patients with Parkinson' s disease in Nanjing city / 中华神经科杂志

Objective To determine the frequency of depression in patients with Parkinson' s disease(PD) and healthy controls over 50 years of age and to analyze the characteristics and influencing factors of PD with depression(PDD) in Nanjing.Methods One hundred and twenty-six PD patients were diagnosed and assessed using Self-rating Depression Scale (SDS) and Hamilton Depression Scale (HAMD).The frequency,characteristics and influencing factors of depression were statistically analyzed,and the factor analysis of HAMD was carried out.Also,one hundred and twenty-four healthy subjects over the age of 50 were selected as the control group.Ressults The incidence of depression in PD group was 48.4％ ( 61/126):15.1％ (19/126) for mild depression,27.8％ (35/126) for moderate depression,5.6％ (7/126) for severe depression.The incidence of depression in the control group was 9.7％ (12/124):5.7％ (7/124) for mild depression,2.4％ (3/124) for moderate depression,1.6％ (2/124) for severe depression.There was a significant difference between these two groups( x2 =45.36,P ＜ 0.01 ).Univariate and Logistic regression analysis revealed that a high frequency of depression occurred in patients with long PD duration,high H-Y stage and UPDRS Ⅲ.According to each factor analysis of HAMD,the scores of cognitive impairment,tardiness,anxiety and sleep disturbances of the PD patients with depressive syndromes were higher than that of the control group.Conclusion Depression is a relatively common complication of PD in Nanjing which is associated with long PD duration,severity of motor disturbances and increasing H-Y stage.

Influence of serum complement and IgG on rituximab-dependent NK cell-mediated cytotoxicity to Raji cells / 白血病·淋巴瘤

Objective To determine the influence of serum complement and IgG on rituximabdependent NK cell-mediated cytotoxicity to Raji cells in vitro.Methods FcγR Ⅲ a (CD16a) polymorphism of NK cells were detected by nest-PCR. Effects of serum IgG on FcγRⅢ a expression of NK cells in vitro were analyzed by flow cytometry.The target cells(Raji cells) were stained with DIO,cultured with effector cells(NK cells) and rituximab with or without serum IgG/complement,and finally stained with propidium iodide (PI),then these cells were tested by flow cytometry and the cytotoxic index was calculated as well. Results The cytotoxic indexes of the ADCC +CDC groups were higher than those of ADCC groups, but the serum IgG groups were lower than the ADCC groups. In FcγRⅢa-158Ⅴ/Ⅴ groups, the cytotoxic indexs of the ADCC+ CDC groups,the serum IgG groups and the ADCC groups were (94.25±1.79) ％,(59.79±0.66) ％ and(69.05± 2.38) ％,respectively,and the differences among the groups were statistically significant (P＜ 0.05).In FcγRⅢ a-158Ⅴ/F groups,the cytotoxic indexs of these three groups were (66.71±5.57) ％,(18.13±2.99) ％ and (39.63±3.86) ％, respectively, and the differences among the groups were also statistically significant (P＜ 0.05).Conclusions Complement may enhance the rituximab-mediated NK cell cytotoxicity to Raji cells, whereas,serum IgG may weaken the cytotoxicity against Raji cells. It is clued up that for patients treated by tumorspecific monolonal antibody (MAb), combined infusion of fresh frozen plasma could promote its anti-tumor effect,however,MAb combined with IVIG may impair its anti-tumor effect.

Association analysis of tryptophan hydroxylase-2 gene polymorphism with antisocial personality disorder and impulsivity / 中华行为医学与脑科学杂志

Objective To study the association of tryptophan hydroxylase-2 (TPH2) gene polymorphism and antisocial personality disorder (ASPD) and its impulsivity in Chinese Han population. Methods The single nucleotide polymorphism (SNPs) of TPH2 in transcriptional control region,-703G/T,was analyzed by PCR-RFLP genotyping assay in 117 ASPD patients and 142 healthy controls. Barratt Impulsiveness Scale-11 (BIS-11) was used to evaluate the impulsivity of subjects. Results There were significant differences between ASPD and controis on genotype and allele frequencies of TPH2-703G/T (x2 = 7.73, P ＜ 0.05; x2 = 5.12, P ＜ 0.05). The GG genotype and G allele were positively associated with ASPD(OR = 1.458,95% CI = 1.080 ～ 1.968 ;OR = 1.479,95% CI = 1.045 ～ 2.094). The scores of BIS-11 and its factors in GG genotype group((71.28 ± 7.50), (19.60 ±3.41), (25.73 ± 4.92), (25.95 ± 4.77) ) were higher than GT genotype group (( 66.23 ± 8.06), (17.79 ±3.02) ,(23.06 ±3.84) ,(25.38 ±4.97)) and TT genotype group((66.55 ±8.49),(18.50 ±3.35),(23.45 ±4.08), (24.97 ± 4.90)), but only the difference of BIS-11 total scores, the attention and motor factor scores among three groups were statistically significant (P＜0.05). The scores of BIS-11 and its factors in G allele group ((69.38 ±8.04), (18.92 ± 3.36), (24.73 ±4. 69), (25.73 ±4.82)) were higher than T genotype group ((66.41 ±8.22),(17.98 ±3.26),(23.27 ±3.94), (25.15 ±4.89)),however,only the difference of BIS-11 total scores, the attention and motor factor scores between two groups were statistically significant.Conclusion TPH2-703G/T polymorphism may be association with ASPD in Chinese Han population. The GG genotype and G allele may be the risk factors of ASPD and impulsivity.

Effects of Mesenchymal Stem Cells Feeder Layer, Culture Sera and Freeze-thaw Lysates on Expansion and Differentiation of Cord Blood CD34~+ Cells in Vitro / 中山大学学报(医学科学版)

[Objective]To investigate the effects of mesenchymal stem cells (MSC) feeder layer, culture sere and freeze-thaw lysates on expansion and differentiation of cord blood CD34~+ cells in vitro. [Methods] MSC were isolated from human bone marrow and cultured until the third passage. Sera were obtained from the cultured MSC, and freeze-thaw lysates were obtained by repeated freeze-thaw procedures. Cord blood CD34~+ cells were isolated by magnetic cell separation system, and were co-cultured with the MSC feeder layer, culture sera, freeze-thaw lysates and hematopeietic growth factors (HGFs), respectively. The nucleated cells, CD34~+ cells, CD34~+CD38~- cells, CD41~+ cells and CD3~+ cells in the above culture system were detected by flow cytometry on day 6 and day 12. [Results] ①MSC feeder layer had a strong effect on nucleated cells, CD34~+,CD34~+CD38~- cells expansion. The MSC sera and freeze-thaw lysates had similar effect on cell expansion, but the effect was weaker than that of feeder layer (P<0.05). ② Both MSC sera and feeder layer inhibited cord blood CD34~+ cells differentiation toward CD3~+ cells or CD19~+ cells, and no significant differences were found between these two groups (P>0.05). ③ Both MSC sera and feeder layer promoted cord blood CD34~+ cells differentiation toward CD41~+ cells, and the effect was stronger in the feeder layer than that of the sera (P<0.05). ④ Freeze-thaw lysates had no effect on cell expansion and differentiation, and were similar with that of HGFs (P>0.05). [Conclusions] The MSC sera have positive effects on expansion of cord blood CD34~+ and CD34~+CD38~- cells, moreover they have the ability of promoting cord blood CD34~+ cells differentiation toward CD41~+ cells.

Osteogenic Activity of MSC Infected by Recombinant Adenovirus Vector Ad-LMP-1 / 中山大学学报(医学科学版)

[Objective]This study was designed to construct a recombinant adenovirus vector contains LMP-1 gene,and investigate the osteoinductive activity of MSC which were transfected recombinated adenoviral vector carrying LMP-1 gene.[Methods]Total RNA was extracted from mt osteoblast and the LMP-1 gene was acquired by RT-PCR,the LMP-1 gene and entry vector pENTR/D-TOPO were used to create the entry clone with the directional TOPO clone technology,then the entry clone and the expression vector were used to create the expression clone throush the LR recombination reaction.The adenovirus expression clone was linearized by PacI and transfected to the 293A cell line to harvest a high titer.Ad-LMP-1 was infected into the 3rd passage MSC,the expression of LMP-1 was detected by Western blot.The osteogenic activity of MSC was evaluated by the expression of collagen Ⅰ,ALP,osteocalcin and the formation of bone nodule.[Result]The LMP-1 gene was successfully acquired and confirmed,the entry clone and the expression clone were both verified by enzymes digestion,and the expression clone was further confirmed by sequenced.The expression of LMP-1 was detected successfully in MSC.The increasing expression of collagen Ⅰ,osteocalcin.ALP and bone nodule were observed by comparing to the control group.[Conclusion]Gateway technology not only make construction of the pAd-LMP-1 recombination adenovirus vector simple and fast,but also get a high transfection efficacy in MSC.LMP-1 gene can induce the osteoblast differention of MSCs,and improve its osteogenic activity.The adenovirus vector is reliable to be used in further gene therapy research.

Exploratory Research of Setting an Optimal Minor-factor Culture System of Cytokine-induced Killer Cells/ Natural Killer Cells / 中山大学学报(医学科学版)

[Objective] In order to explore the optimal minor-factor culture system of CIK/NK cells by proliferating CIK/NK cells using single-factor,bi-factor,and tri-factor combinations of IL-2,IL-7,and IL-12.[Methods] Ficoll-Hypaque method was used to separate cord blood mononuclear cells and divide them into 6 groups.Add single-factor,bi-factor,and tri-factor combinations (IL-7;IL-12;IL-7 + IL-12;IL-2 + IL-7;IL-2 + IL-12;IL-2 + IL-7 + IL-12) of IL-2 (80 ng/mL),IL-7 (40 ng/mL),and IL-12 (40 ng/mL) and culture them in total 21 days then harvest the cells.To collect cell suspensions of each group in culture outset and day 21 and to count the proportion of CD3+ CD56+ CIK cells and CD3- CD56+ NK cells with flow cytometry.[Results] The proportion of CIK cells in single-factor culture system of IL-12 was the highest in all groups (30.23 ± 1.18%).The proportion of CIK cells in bi-factor culture system of IL-2 + IL-12 can raised to 18.58 ± 0.68%.The proportion of CIK cells of the two groups above can reach the level of that using traditional multi-factor culture methods.NK cell proportion in IL-12 was 30.23 ± 1.18%,NK cell proportion in IL-2 + IL-7 can also reach to 29.52 ± 0.89%.[Conclusions] It is adoptable to proliferate CIK/NK cells using minor-factor culture system of IL-12,IL-2 + IL-12,or IL-2 + IL-7.

Teratogenecity of Ethylene Oxide Sterilized Medical Devices on Cells Used for Biotherapy / 中华医院感染学杂志

OBJECTIVE To observe the teratogenecity of ethylene oxide sterilized materials to cells which are used for biotherapy.METHODS Three kinds of cells were cultivated in ethylene oxide sterilized culture flasks which were conserved for different time and high temperature/high pressure sterilized vitric culture flasks.NK/CIK cells induced from cord blood,human mesenchymal stem cell and CD34+ cells were cultivated in ethylene oxide sterilized culture flasks which were conserved for one day(group A),for three months(group B),for more than six months(group C) and cultivated in high temperature/high pressure sterilized vitric culture flasks(group D).After ten days,not only the growth and morphology of cells but also the breakages fragmentations and gaps in chromosomes and replication of chromosome in nucleus were observed.According to these data,the malformation rate was calculated.RESULTS Compared with groups C and D,the inductivity of groups A and B obviously degraded(P0.05).CONCLUSIONS Compared with the control group,the materials which were sterilized by ethylene oxide and conserved for less than one day and three months can bring damage to cells.However,the damage is not discovered in the materials which were conserved for more than six months.Ethylene oxide can induce cell aberrant and mutation.Therefore,it is suggests that materials be conserved for more than six months to ensure safe and effective enforcement of biotherapy.

Thalassemic serum panel reaction antibody inhibits proliferation and differentiation of cord blood hematopoetic stem cells in children patients / 中国组织工程研究

BACKGROUND: Panel reaction antibody (PRA) plays an important role in rejection of recipients undergoing solid organ transplantation, which has a positive effect on nonfunction of implant. OBJECTIVE: To evaluate the effect of thalassemic serum-specific PRA on the proliferation and differentiation of umbilical cord blood hernatopoetic stem/progenitor cells (HSC/HPCs) in children patients with thalassemia. DESIGN, TIME AND SETTING: The in vitro cytology experiment was performed at the Experimental Research Center, Second Affiliated Hospital, Zhongshan University from January 2006 to August 2007. MATERIALS: Five samples of umbilical cord blood from healthy full-term birth puerperants (each 80 100 mL) were used in this study. PRA serum samples of children patients with thalassemia after repetitive blood transfusion, five samples of AB blood grouping serum, and six samples of positive anticoagulation vein blood (10 mL) were used in the study. METHODS: Mononuclear cells were harvested from umbilical cord blood by Ficoll-Hypaque gradient centrifugation. 1 × 105 rnononuclear cells from umbilical cord blood were incubated with different levels of experimental or AB control serum (0, 50, 100 μ L) from healthy children. The mixture mentioned above was incubated with rabbit complement for semisolid colony culture.MAIN OUTCOME MEASURES: Colony-forming units (CFU) were counted and observed after 7 days and 14 days of culture under an inverted microscope.RESULTS: After incubation with HSC/HPCs PRA serum, total number of CFUs and varied CFUs decreased to different extents, of which the total number of CFUs and CFU- granulocyte-rnacrophages (CFU-GM) had significant differences (P < 0.01). Moreover, there were negative correlations between different levels of serum PRA and the followings: number of total colonies, CFU- GM, CFU- granulocyte-erythrocyte-monocyte-megakaryocytes, CFU-erythroids, burst forming unit-megakaryocytes, and CFU-megakaryocytes (P < 0.05).CONCLUSION: The thalassemic serum PRA has an apparent inhibitory effect on the proliferation and differentiation of cord blood HSC/HPCs in vitro, an effect that may be pronounced with increasing serum PRA.

Delayed transplantation of olfactory ensheathing cells for thoracic cord injury in adult rats / 中国组织工程研究

BACKGROUND: Spinal cord can regenerate after injury in certain microenvironment. Olfactory ensheathing cells (OECs)have the characteristics of astrocytes and Schwann cells and can accelerate the spinal cord axonal regeneration.OBJECTIVE: To make injured thoracic cord rat models and observe the effect of OECs on injured spinal cord axonal regeneration.DESIGN: Observational experiment.SETTING: Second Affiliated Hospital of Sun Yat-Sen University.MATERIALS: The experiment was performed at the Second Affiliated Hospital of Sun Yat-Sen University from January 2001 to November 2002.Totally 20 adult SD male rats with the body mass of (380±20) g were provided by Experimental Animal Center of Sun Yat-Sen University (number of institution license SYXK2004-0020). There were DMEM culture solution with low glucose (L-DMEM, GibcoBRL), fetal calf serum (FCS) (Hyclone), myelin basic protein (MBP) (Sigma) and nerve growth factor receptor antibody (Sigma). They were divided into cell transplantation group and control group by the method of random digits table with 10 in each group.METHODS: The adult SD rats were anaesthetized and decapitated to obtain the whole olfactory bulb and then isolate olfactory nerve with a sterile operation. Thoracic cord injury models were established by modified Allen method. 10μL OECs suspension (2.5×1010 L-1) was injected into injured spinal cord of the cell transplantation group, whereas DMEM/F12 (1:1) culture solution of the same dose was injected in the control group. The influence of OECs on spinal cord axonal regeneration was observed by histological and immunohistochemical method 6 weeks after transplantation.MAIN OUTCOME MEASURES: ①OECs were identified by nerve growth factor receptor antibody staining. ②Repair of myelin sheath was observed by MBP staining. ③Nerve axonal regeneration was observed by argentaffin staining. RESULTS: Two rats in the cell transplantation group and 3 rats in the control group died, so totally 15 rats were involved in the result analysis. ①In the cell transplantation group,injured spinal meninges were integrated,but spinal cord became thin as compared with the normal spinal cord. By hematoxylin-eosin (HE) staining, multipolar cells appeared in injured region and the cells were fused excessively with myeloid tissues. It proved that survival OECs were integrated well within the host. New axons were clustered in bundles and infiltrated by small round lymphocytes. By argentaffin staining, regenerated axons grew in tissues of injured region, which mostly accompanied with fascicular-arranged multipolar cells. In the control group, spinal cord became thin markedly and spinal meninges were integrated. No new axon appeared in the injured spinal cord by HE staining. No regenerated axon appeared by argentaffin staining, either. ②In the cell transplantation group, most multipolar cells were clustered in bundles. A mass of positive granules of nerve growth factor receptor antibody appeared in cytoplasm, which further verified that OECs still survived and integrated well within the host 6 weeks after transplantation. Linear MBP positive fibers appeared in the injured region by MBP staining,which indicated that myelin-like substance appeared and drew closely in both ends of injured region. At the same time,MBP positive substance also appeared in the multipolar cells, which illustrated that transplanted OECs could induce the occurrence of myelin-like substance. No regenerated axon was found in the control group.CONCLUSION: Delayed transplantation of OECs can survive and induce the occurrence of myelin-like substance in injured spinal cord of adult rats.

Biological characteristics of olfactory ensheathing cells after transplantation into spine cord / 中国组织工程研究

BACKGROUND:Olfactory ensheathing cells (OECs) have been shown to possess the potential of repairing injured spinal cord, but their biological characteristics after transplantation in vivo are not well understood.OBJECTIVE: To investigate the migration of OECs after transplantation into the injured spinal cord of adult rats.DESIGN: Randomized and controlled experiment.SETTING: Department of Orthopedics, Guangdong Provincial People's Hospital; Experimental Animal Center of the North Campus of Sun Yat-sen UniversityMATERIALS: Totally 38 2-month-old male SD rats with body mass of (350 ±20) g were used in this study.METHODS: This experiment was conducted in the Experimental Animal Center, North Campus of Sun Yat-sen University between February 2004and May 2004. Two SD rats were used to extract the OECs, which were stained with Hoechst 33342. Totally 36 SD rats were subsequently randomized into 3 groups, namely rostral transplantation group, caudal transplantation group and control group with 12 rats in each group. The rats in the rostral and caudal transplantation groups subjected to operations to establish thoracic spinal cord injury model and OEC suspension was injected; in the control group, the rats were spared of thoracic spinal cord injury with only OEC suspension injection.MAIN OUTCOME MEASURES: Distribution of OECs in the spinal cord was observed under fluorescence microscope 1, 2, 4 and 6 weeks after operation, respectively.RESULTS: Of the rats in the 3 groups, 1 died in the rostral group, and 2in each of the caudal transplantation and control groups, leaving 29 rats for result analysis. The OECs in the rostral and caudal transplantation groups migrated longitudinally along the long axis of the spinal cord to a farthest distance of 8 mm and penetrating the scar tissues, but very few cells could reach the contralateral side. The OECs in the control group diffused locally without migration.CONCLUSION: OECs mainly migrate along the axons in white matter of the injured spinal cord, and their rostral and caudal migration does not differ in speed or amount. Only a small amout of OECs can across the transected gap of the spinal cord.

Influences of olfactory ensheathing cells transplantation on axonal regeneration in spinal cord of adult rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe whether olfactory ensheathing cells could be used to promote axonal regeneration in a spontaneously nonregenerating system.</p><p><b>METHODS</b>After laminectomy at the lower thoracic level, the spinal cords of adult rats were exposed and completely transected at T10. A suspension of ensheathing cells was injected into the lesion site in 12 adult rats, and control D/F-12 (1:1 mixture of DMEM and Ham's F-12) was injected in 12 adult rats. Six weeks and ten weeks after cell transplantation, the rats were evaluated by climbing test and motor evoked potentials (MEPs) monitoring. The samples were procured and studied with histologicl and immunohistochemical methods.</p><p><b>RESULTS</b>At the 6th week after cell transplantation, all the rats in both the transplanted and control groups were paraplegic and the MEPs could not be recorded. At the 10th week after cell transplantation, of 7 rats in the control group, 2 rats had muscles' contraction of the lower extremities, 2 rats had hips and/or knees' active movement; and 5 rats' MEPs could be recorded in the hind limbs in the transplanted group (n=7). None of the rats in the control group had functional improvement and no MEPs recorded (n=7). Numerous regenerating axons were observed through the transplantation and continued to regenerate into the denervated host tract. Cell labelling using anti-Myelin Basic Protein (MBP) and anti-Nerve Growth Factor Receptor (anti-NGFR) indicated that the regenerated axons were derived from the appropriate neuronal source and that donor cells migrated into the denervated host tract. But axonal degeneration existed and regenerating axons were not observed within the spinal cords of the adult rats with only D/F-12 injection.</p><p><b>CONCLUSIONS</b>The axonal regeneration in the transected adult rat spinal cord is possible after ensheathing cells transplantation.</p>