1.In vitro culture of autologous mesenchymal stem cells from the joint drainage fluid after knee arthroscopy: a feasibility study
Xuezhen LIANG ; Bo XU ; Shaoshan WANG
Chinese Journal of Tissue Engineering Research 2017;21(21):3306-3311
BACKGROUND:Mesenchymal stem cells have a extreme prospect in orthopedics, which show great potential especially in the treatment of articular cartilage defect disease. Bone marrow is the main source of mesenchymal stem cells, and the iliac puncture is a conventional way to obtain bone marrow, but is restricted by the limited resources and strict technical requirements. Therefore, it is of great significance to explore new effective and convenient sources of mesenchymal stem cells. OBJECTIVE:To explore the feasibility of autologous mesenchymal stem cells derived from the joint drainage fluid after knee arthroscopy.METHODS: We selected eight patients who underwent arthroscopic surgery to collect joint drainage fluid by pre-made sterile blood bag before the wound closure. Precipitation with hydroxyethyl starch and density gradient centrifugation method were performed to isolate and culture mesenchymal stem cells from the joint drainage fluid. Cell morphology, growth curve, surface marker identification were observed and detected using flow cytometry. Then, adipogenic, chondrogenic and osteogenic differentiation of cells were induced and identified by oil red O, toluidine blue staining, and alizarin red staining, respectively. RESULTS AND CONCLUSION:The cultured cells were spindle-shaped, adherently grew and had good proliferation ability, which were positive for CD44, CD90, CD105 and CD73, but not for CD45. Under standard inductions, the cultured cells were induced to differentiate into osteoblasts, adipocytes and chondrocytes. Therefore, these cells were confirmed as mesenchymal stem cells. Mesenchymal stem cells were successfully isolated from the joint drainage fluid of eight patients and had no difference in cell morphology, proliferation and phenotypes. To conclude, the joint drainage fluid is an ideal source of mesenchymal stem cells with the guaranteed quality and quantity.
2.Detection of clostridium perfringens with qPCR in mouse models and a clinical case
Yuling SHI ; Shaoshan XU ; Zhaohui SUN ; Lidan CHEN ; Lingling TANG
Journal of Chinese Physician 2011;13(3):324-327
Objective To detect of clostridium perfringens by qPCR in mouse models and a clinical case in order to offer early diagnosis.Methods 40 Kunming mice were randomly grouped and intramuscular injected clostridium perfringens type A in leg 0.1 ml(3.5 × 109cfu/ml or 3.5 × 108cfu/ml or 3.5× 107cfu/ml,diluted with saline),while control group was injected with 9% sodium chloride 0.1ml.The mouse models and a clinical case were detected by qPCR.Results The death rate of 3.5 × 109,3.5 × 108,3.5 × 107cfu/ml and the blank group were 90%,70%,10% and 0% after intramuscular injection for 72 h spectively.The mean Ct values among these groups were 21.21 ±2.69,28.45 ±2.74,32.49 ±2.87 and 0.00 ± 0.00(P < 0.05).The Ct values of the patient were 30.67 and 30.44.Conclusions Cclostridium perfringens could be successful identified with qPCR in mouse models when the mice still did not show any symptoms.
3.Effect of α-phenyl-N-tert-butylinitrone on Expression of Nerve Growth Factor in Spinal Cord Injured Rats
Shaohui LIU ; Wei LIU ; Shaoshan QIU ; Xijin XU
Chinese Journal of Rehabilitation Theory and Practice 2014;(8):709-712
Objective To explore the effect of α-phenyl-N-tert-butylinitrone (PBN) on expression of nerve growth factor (NGF) in serum and spinal cord tissue in rats after spinal cord injury (SCI). Methods 174 female Sprague-Dawley rats were randomly assigned to following groups: normal control group (n=54), normal saline control group (NS group, n=60, intrathecally injected normal saline 15 μl), and PBN group (n=60, intrathecally injected PBN, 3 mg, 15 μl). The model was established with New York University blow device (150 kDyne, 1 s dwell time). PBN was intrathecally injected into the damaged areas 30 min after operation, then once a day for 7 days. The Basso-Beattie-Bresnahan (BBB) Locomotor Rating Scale was used to assess the rats 3 days and 1 day before, and 1 day, 5 days, 10 days, 15 days, 20 days,25 days, 30 days and 35 days after SCI. NGF in the injured spinal cord tissue and serum was measured 1 h, 12 h, 24 h, 48 h, 3 days, 7 days,14 days and 21 days after SCI. Results NGF increased in serum but not in spinal cord. The ratio of NGF/total protein in serum rose and peaked 48 h after SCI, and the ratio was higher in NS group (0.92%±0.02%) than in PBN group (0.77%±0.05%) (P=0.021). BBB scores increased from the 9th day, and PBN group improved better than NS group (P<0.01). Conclusion PBN could reduce the expression of NGF in the SCI rats, and promote the recovery of neurol function.
4.Pathological analysis of time-zero renal biopsy in donor kidney
Ping LI ; Yiyao TU ; Shaoshan LIANG ; Feng XU ; Dandan LIANG ; Jingsong CHEN ; Zhihong LIU ; Caihong ZENG
Chinese Journal of Organ Transplantation 2019;40(6):355-360
Objective To summarize the pathological survey of time-zero renal biopsy (T0-RBx ) . Methods The material qualities and pathological features were analyzed retrospectively for T 0-RBx (n=176) between March 2008 and May 2016 .According to the source of donor kidney ,T0-RBx specimens were divided into living donors (LD) group (n=137) and Deceased donation (DD) group (n=39) .Furthermore , the DD group was divided into cerebral hemorrhage group (n= 10) and brain trauma group (n= 29) according to the causes of death .The inter-group differences of pathological characteristics and the effects of abnormal pathological lesions on allograft function were observed .Results All T0-RBx specimens contained cortical kidney tissue .The average microscopic length of renal tissue was (0 .39 ± 0 .23) cm and the median glomerular number 11 . The abnormal pathological lesions included glomerulosclerosis (GS ,30 .7 % ) , segmental glomerulosclerosis (1 .1 % ) ,mesangial increase (MI ,19 .3 % ) ,tubular atrophy (TA ,35 .2 % ) , acute tubular necrosis (ATN ,9 .1 % ) ,vacuolar degeneration of tubular epithelium (27 .3 % ) ,losses in tubule epithelial brush border (97 .7 % ) , protein cast (25 % ) , interstitial fibrosis (IF ,34 .1 % ) , inflammation (I ,42 .6 % ) ,arteriolar hyalinosis (AH) (26 .1 % ) and vascular fibrous intimal thickening (CV ,23 .3 % ) .Among them ,23 .9 % ,1 .1 % ,0 .55 % and 0 .55 % cases were diagnosed as IgA nephropathy ,immune complex associated with glomerular disease and focal segmental glomerulosclerosis diabetic nephropathy respectively .And the reminders were of ischemic injury .The incidence rates of TA ,IF and I were lower in DD group than those in LD group ( P< 0 .05 ) . However , ATN and vacuolar degeneration of tubular epithelium were higher (P<0 .001) .The incidence of GS was significantly higher in cerebral hemorrhage group than that in brain trauma group (P<0 .01) .No statistical difference existed in other lesions or disease constitution among the groups (P>0 .05) .Further analysis showed GS was related with allograft function at 6/12 months post-transplantation in both LD and DD groups (P<0 .05) .IF and AH were also related to short-term renal function of recipients post-transplantation in LD and DD groups (P>0 .05) .Conclusions T0-RBx may detect the abnormal lesions of donor kidney .Some differences exist in types and degree of abnormal lesions among different donor kidneys .LD group has a higher risk for chronic histological injury such as TA and IF while DD group is more susceptible to acute renal tubular interstitial injury .Thus it is valuable for predicting allograft function post-transplantation .Material quality is essential for ensuring the reliability of T 0-RBx .
5.Evaluation of effect of oral health intervention on children in Shaoshan area of Hunan province.
Zi Yi ZHANG ; Bin XIA ; Ming Ming XU ; Yi Ping LI ; Zhan Gui TANG ; Yong Qing CHEN
Journal of Peking University(Health Sciences) 2020;52(5):913-918
OBJECTIVE:
To understand the oral health status of children aged 3-12 in Shaoshan area of Hunan province and to evaluate the role of oral health educations based on community such as fluoride varnish, oral hygiene introduction in improving the oral health of children in the area so as to gain expe-rience.
METHODS:
The study used cluster sampling to select 3 kindergartens and 2 primary schools in different economic development areas of Shaoshan. Oral health status survey and interventions were conducted in December 2014 and September 2016, respectively. The average debris index, decayed missing filled teeth (deciduous teeth: dmft; permanent teeth: DMFT), and caries prevalence rate of children aged 3-6 years and 6-12 years were compared. At the same time, children aged 5 and 12 were used as representative populations to compare the indices as listed before and the caries prevalence rate of the first permanent molar in 12-year-old children was compared as well. Finally, health economic analysis was carried out based on the 2 years' result.
RESULTS:
In this study, 992 children and 896 children in 2014 and 2016 were included respectively. As for children aged 3-6 years, the average debris index and dmft in 2016 were significantly less than that in 2014 (P < 0.001, P < 0.001), and the difference of DMFT was not significant (P=0.419). Children aged 6-12 years showed the same result, the average debris index and dmft in 2016 were significantly less than those in 2014 (P < 0.001, P=0.013), and the difference in DMFT was not significant (P=0.674). 173 and 179 5-year-old children were included in 2014 and 2016 respectively, and the dmft showed significant difference (P=0.038); the caries prevalence rate was 75.7% and 71.5%, respectively, which was also not significant (P=0.370). With respect to the 12-year-old children, 65 and 104 children were included and the differences in dmft and DMFT were not significant (P=0.133, P=0.171). The caries prevalence of the first permanent molar in the 12-year-old children was 36.9% and 26.9%, whose difference was not significant (P=0.171).
CONCLUSION
The application of fluoride varnish and oral health education can significantly reduce the dmft of children aged 3-12 years in Shaoshan area and significantly improve the oral hygiene status. DMFT, the caries prevalence rate of 5-year-old children's deciduous teeth and 12-year-old children's first permanent molar showed a decline.
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