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.Application of 1H-MR spectroscopy in crerbral glioma
Shuang LIANG ; Shaoshan LI ; Qiang FU ; Qingjiu ZHOU ; Bo LIU ; Chen LIU
Chinese Journal of Nervous and Mental Diseases 2015;(9):552-557
Objective To investigate the application of 1H-MR spectroscopy in predicting the value of each grade range and grading of glioma invasion. Methods A restrospective analysis was conducted on the preoperative biopsy or posoperative pathology confirmed glioma in 33 cases. Cerebral gliomas were graded into four categories based on the stan?dard of WHO(2007):grade I, II, III andⅣ. All patients underwent MRI and 1H-MR spectroscopy before operation. The 1H-MR spectroscopy analysis was used to analysis metabolite NAA,Cho,Cr and NAA/Cho, NAA/Cr, Cho/Cr in the tu?mor area, peritumoral region and normal area. Results 1H-MRS revealed that there were no significantly differences in Cho, Cho/Cr, NAA/Cho between the peritumoral region and tumor area of grade I glioma (P<0.025). However, there were significantly differences in Cho, Cr, NAA/Cho, Cho/Cr ratio between peritumoral region and the normal brain tissue of grade I glioma (P<0.025). There were significantly differences in NAA, NAA/Cho, NAA/Cr ratio between peritumoral re? gion and the tumor area of grade II glioma (P<0.025). There were statistically significantly differences in Cho, NAA, Cr, NAA/Cho, NAA/Cr, Cho/Cr ratio between the peritumoral region and normal brain tissue of Grade III orⅣglioma (P<0.025). The ratio of Cho/Cr and NAA/Cho were correlated with the pathological grade(F values were 403.9 and 159.46, P<0.05), and the ratio of NAA/Cr and NAA/Cho were decreased gradually whereas the ratio of Cho/Cr was gradually in?creased(F=119.91,P<0.05). Conclusion The 1H-MR spectroscopy analysis has a predictive value in evaluating the lev?el of glioma invasion and determining the grading of glioma before surgery, which is very helpful for the surgical plan and the scope of resection, thereby reducing the recurrence and prognosis of the patients after surgery.
3.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 .
4.Short-term prognosis of kidney allograft evaluated by pre-implantation biopsy combined with Lifeport
Xue LI ; Shaoshan LIANG ; Dongrui CHENG ; Jiqiu WEN ; Kenan XIE ; Xuefeng NI ; Jinsong CHEN
Chinese Journal of Organ Transplantation 2021;42(5):287-292
Objective:We aimed to evaluate the predictive value of pre-implantation biopsy combined with Lifeport for the short-term prognosis of kidney allograft from donation after citizen death (DCD).Methods:Data from a total of 34 patients who had undergone kidney transplantation in Jinling Hospital from December 2017 to December 2019 were retrospectively analyzed. Histopathological data from pre-implantation biopsy , Lifeport parameters and recipient kidney transplant function at 3 months post-surgery were collected. The performances of histopathological indexes , and Lifeport parameters to predict delayed graft function (DGF) and estimated glomerular filtration rate (eGFR) at 3 months post-surgery were observed evaluated.Results:13 cases of DGF occurred, accounting for 38.2%. Serum creatinine at death and resistance index (RI) at 0.5 h, 1 h, 2 h and 4 h after Lifeport hypothermic machine perfusion (HMP) in the DGF group was significantly higher than that in the non-DGF group. Histologically, the acute tubular injury (ATI) score of the DGF group was higher than that of the non-DGF group, whereas the Remuzzi score was not statistically different between the two groups. The eGFR at 3 months post-transplant was moderately correlated with the RI at 4 h HMP and the Remuzzi score (RI: r=-0.48, P<0.001; Remuzzi score: ρ=-0.42, P=0.01), but no correlated with ATI score of the donor kidney. Although Remuzzi score was not correlated with kidney allograft recovery time (ρ=-0.25, P=0.16), it was inversely correlated with eGFR at 3 months post-transplant (ρ=-0.42, P=0.01). Combined use of Lifeport HMP 4-hour RI and ATI score increased the sensitivity and specificity of predicting DGF to 100% (95% CI: 75.3%-100%) and 90.5% (95% CI: 69.6%-98.8%) respectively. Conclusions:The serum creatinine at death, Lifeport RI, and ATI score of the DGF group were significantly higher than those of the non-DGF group, and the eGFR at 3 months post-transplant was correlated with the Lifeport RI and Remuzzi score. Combined use of ATI score and RI at 4 hours of Lifeport perfusion improved the sensitivity and specificity of predicting DGF .