Asian Continental Ancestry Group ; Calcinosis ; Colitis ; pathology ; Female ; Humans ; Middle Aged

BACKGROUND:We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cels at-80℃.
OBJECTIVE:To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cels and to verify the biological features of bone marrow mesenchymal stem cels after long-term cryopreservation.
METHODS: Bone marrow mesenchymal stem cels were cultured using adherent method and the biological features and purity of cels were detected using immunofluorescence method. Bone marrow mesenchymal stem cels were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at-80℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cels. After 1, 3, 6 months of cryopreservation, the cels were resuscitated, cultured and passaged. Passage cels were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cels cryopreserved at-80℃.
RESULTS AND CONCLUSION:Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80℃, and resuscitated cels were able to proliferate in vitro, and passage normaly, indicating the cryopreserved bone marrow mesenchymal stem cels stil maintain the original biological activity.

Objective To evaluate the feasibility and safety of hypothermic perfusion via renal artery balloon catheter before laparoscopic partial nephrectomy.Methods A total of 35 patients,who were arranged to receive laparoscopic partial nephrectomy during the period from March 2013 to December 2016 at the General Hospital of PLA,China,were enrolled in this study.The tumor was located in the left kidney in 22 patients and in the right kidney in 13 patients.The long diameter of the tumors was 2.3-7.0 cm,with a mean of (4.1±1.2) cm.Before laparoscopic partial nephrectomy,the implantation of renal artery balloon catheter was performed in all patients.The intraoperative renal cold-ischemia time,the time spent for operation and the amount of intraoperative blood loss were recorded.The creatinine clearance rate (Ccr) and estimated glomerular filtration rate (eGFR) were determined before operation as well as at one,3 and 7 days after operation.The intraoperative and postoperative complications were analyzed.Results Successful implantation of renal artery balloon catheter was accomplished in all 35 patients,with the technical success rate being 100％.Slipping of balloon catheter before laparoscopic partial nephrectomy occurred in one patient;in 2 patients incomplete occlusion of renal artery was found during operation and artery forceps had to be used to occlude the renal artery.In the other 32 patients,laparoscopic partial nephrectomy under the condition of hypothermic perfusion via renal artery balloon catheter was successfully carried out.No severe complications occurred during and after the operation.The average cold-ischemia time of the diseased kidney was 45 (20-125) min,the mean time for interventional procedure was 28 (20-40) min,the average surgery time was 147(95-235) min,the average amount of intraoperative blood loss was 180(50-1000) ml.The Ccr values determined before and at one,3,7 days after nephrectomy were (96.5±15.1),(75.2±10.5),(54.3±13.8) and (91.8±14.1) ml/min respectively.The eGFR values determined before and at one,3,7 days after nephrectomy were (99.5±15.3),(70.3±12.5),(65.5±11.7) and (96.8±12.3) ml·min-1·m-2 respectively.Statistical analysis indicated that in both groups the preoperative and 7-day postoperative Ccr values and eGFR values were significantly different from their one-day and 3-day postoperative values (P＜0.01),but no statistically significant differences existed between preoperative values and 7-day postoperative ones (P＞0.05).Conclusion Hypothermic perfusion via renal artery balloon catheter before laparoscopic partial nephrectomy is clinically safe and feasible,it can prolong the safe time of renal ischemia and protect renal function.

Objective To evaluate the efficacy and safety of acute ureteroscopy for the treatment of ureterie stones during middle and late pregnancy.Methods From June 1998 to March 2005,17 pregnant women(mean age,27 years;age range,21-35 years)with ureteric stones were treated by ureteroscopy when the fetus was at 20-36 weeks of gestation(mean,29 weeks).All the cases presented with urgent symptoms such as recurrent renal colic(11 cases),fever(4)or acute obstructive anuria(2).Among 17 cases,the stones(between 6 mm?7 mm and 13 mm?21 mm)were located in the upper(8 cases),middle(5)or lower ureter(4);and on the left side(5 eases),on the right(10)and on both(2)of the lower ureter. Mild hydronephrosis were observed in 6 cases and moderate hydronephrosis in 11,Of the 17 cases,14 under- went ureteroscopic pneumatic lithotripsy;in 1 case the calculi were pushed to the renal pelvis;and 2 cases were treated by Double-J catheter drainage.Results All the urgent symptoms in 17 cases were relieved after treatment.The stone-free rate of initial treatment was 82.4%(14 of 17).Three cases with residual stones were treated by Douhle-J catheters,which were replaced every 3 months until the calculi were re- moved.No abortion,premature delivery or complications such as ureter perforation occurred.Mild renal colic occurred in 1 case after insertion of Douhle-J catheter,and it was relieved 3d later;gross hematuria occurred in l case and disappeared 6 d later without treatment.All 17 patients had normal delivery and gave birth to healthy children.Conclusions Ureteroscopy is a safe and reliable method for the treatment of ureteric calculi during middle and late pregnancy.

AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.

Objective To investigate the effect of ursolic acid(UA) on NOX2/ROS/NLRP3 inflammasome activation in carbon tetrachloride(CCl4)-induced liver fibrosis SD rat,and to observe the improvement of collagen deposition in liver tissues. Methods All rats were randomly divided into 3 group:control group,CCl4model group,UA treatment group. Liver fibrosis model SD rats was established by the CCl4-induced method and half of them was used as UA treatment group. Serum ALT was detected by ALT detection kit.The liver pathology and collagen deposition were ob-served by HE and Sirius-red staining. The mRNA expression of Nox2,Nlrp3,Caspase1,IL-1β in liver tissues was detected by RT-qPCR. The protein expression of NOX2,NLRP3,caspase-1 and IL-1β in liver tissues was detected by Western blot and immunohistochemistry and the ROS generation in liver tissues was detected by DCFH-DA fluores-cence probe. Results Compared with control group,in the CCl4model group,the serum ALT was much higher (P<0.05);the Ishak's fibrosis score and collagen deposition was significantly increased(P<0.05) and mRNA of Nox2, Nlrp3,Caspase1,IL-1β was increased.In addition,both the NOX2,NLRP3,caspase-1 p10 and IL-1β protein expres-sion and ROS level (P<0.05) of CCl4model group were significant increased.Compared with CCl4model group,in the UA treatment group Ishak's fibrosis score,collagen deposition and ALT decreased.Both mRNA expression of the Nox2, Nlrp3,Caspase1,IL-1β and protein expression of NOX2,NLRP3,caspase-1 p10 and IL-1β as well as ROS were signif-icant decreased,but the caspase-1 p45 protein level has no difference among all these groups (P>0.05). Conclusions Ursolic acid attenuates the liver injury and reduces the collagen deposition,which may relate to its inhibitory effects on NOX2/ROS/NLRP3 inflammasome activation to reduce IL-1β releasing.

This paper is aimed to report the development of a method for the determination of the binding rate of plasma protein with salvianolic acid B. In vitro, equilibrium dialysis method was used to imitate the binding process between salvianolic acid B and plasma protein, in vivo, ultrafiltration method was used and the binding rate with HPLC was determined. Plasma samples were treated with methanol to precipitate the protein, and the buffer solution was directly determined after filtering. The calibration curve of the buffer solution was linear in the range of 0.5-20 microg mL(-1). The calibration curve of the plasma was linear in the range of 2-200 microg mL(-1). The extract recovery was 68.6%-81.9%. RSDs of intra- and inter-day precisions were all less than 8.5%. The binding rates of plasma protein with salvianolic acid B in vitro was 75.2% and in vivo was 92.1%. This paper shows the high binding power of salvianolic acid B to plasma protein with high sensitivity, good reproduction, simple management and fulfilling the requirement.
Animals ; Benzofurans ; blood ; isolation & purification ; metabolism ; Blood Proteins ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Male ; Protein Binding ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Sensitivity and Specificity ; Ultrafiltration ; methods

OBJECTIVETo evaluate the effects of school entrance age on cognition and behaviors in children with attention deficit hyperactivity disorder (ADHD) using mathematical event-related potential (ERP), behavioral test, and Conners Parent Symptom Questionnaire (PSQ).

METHODSFifty-eight ADHD children aged 7-12 years were enrolled and classified into older age and younger age groups according to the school entrance age (n=29 each). The children in the older age group were admitted at an age of 6 years and 6 months to 6 years and 11 months, and those in the younger age group were admitted at an age of 6 years to 6 years and 5 months. The ERP with a mathematical task was used to detect the difference in brain electrical activity between the two groups, and the behavioral test results were compared. The children's parents were asked to complete the PSQ, and the scores on each subscale were compared.

RESULTSThe ERP detection showed that the older age group had a significantly higher P2 amplitude for wrong answers than the younger age group (10.9±5.0 μv vs 8.5±3.6 μv; P<0.05). The younger age group had a significantly shorter time of response to wrong answers than the older age group (619±340 ms vs 870±418 ms; P<0.05). The scores on the subscales of learning problems and impulse-hyperactivity of PSQ were significantly higher in the younger age group than in the older age group (P<0.05).

CONCLUSIONSSchool entrance age can affect cognition and behaviors in children with ADHD, and the ADHD children with a younger school entrance age have an obvious defect in executive function, especially the function of error detection, which leads to the prominent problems in impulse-hyperactivity and learning.

Age Factors ; Attention Deficit Disorder with Hyperactivity ; physiopathology ; psychology ; Child ; Child Behavior ; Evoked Potentials ; physiology ; Female ; Humans ; Male

Objective To investigate the value of perfusion imaging in predicting the status of isocitrate dehydrogenase 1 (IDH1) in glioblastoma.Methods Retrospectively studied 30 patients with glioblastoma multiforme with wild type IDH1 (IDHw) and 30 patients with mutant IDH1 (IDH1m) in Department of Neurosurgery,Shanxi Provincial People's Hospital from September 2014 to February 2016.Ktrans and Ve within the enhancing portion of each tumor were measured by using DCE-MRI data.rCBF and rCBV within the enhancing portion of each tumor were measured by using DSC-MRI.Four parameters were represented as (x) ± s,each of the 4 parameters was compared between patients with wild type IDH1 and mutant IDH1 by using the t-test.The performance in discriminating between the two entities was evaluated by using receiver operating characteristic analysis.Results Ktrans,Ve,rCBF and rCBV inside the enhancing lesion were significantly higher in patients with wild type IDH1 than in those with mutant IDH1.There was a statistically significant difference between IDH1w group and IDH1m group of rCBF value (P ＜ 0.05).The area under the curve for Ktrans,Ve,rCBF,and rCBV inside the enhancing lesion were 0.850,0.873,0.739 and 0.772,respectively.The value of rCBF value of the Ktrans value Ve value of IDH1w was significantly higher than that of IDH1m(P ＜0.05) in the value of the Ktrans value Ve value of rCBF,the value of the AUC value in rCBV was not significantly different with the combination of the 4 parameters of the diagnostic performance (AUC =0.915).Conclusions Ktrans,Ve,rCBF and rCBV calculated from MRI are useful for predicting the IDH1 mutation status.This method is not only for the classification of brain glioblastoma diagnosis has important value,and in glioblastoma classification is of important value in the preoperative noninvasive evaluation,and it has reference significance for predicting the prognosis of patients.

Objective To explore the effect of silencing focal adhesion kinase (FAK) gene on the proliferation and migration of human tongue squamous cell carcinoma cell lines CAL-27.Methods The siRNA interference technology was used to complete the construction of FAK siRNA by transient transfection.CAL-27 cells were divided into the experiment group,the negative control group and blank control group.The expressions of FAK mRNA and protein were detected by qPCR and Western blotting respectively.The cell proliferation ability was tested by using MTT assay.The cell migration ability was detected by using Transwell chamber assay.Results The expression level of FAK mRNA and protein in experiment group was lower than that in blank control group and negative control group,the difference was significant (PinRNA ＜ 0.01,Pprolein ＜ 0.05).Compared with negative control group and blank control group,the proliferation ability of cells in experiment group was obviously decreased at 48 h and 72 h (P ＜ 0.01).Transwell chamber migration assay showed,the average number of migrating cells in experimnet group group was lower than that in blank control group and negative control group,the difference was significant (P ＜ 0.05).Conclusions FAK gene silencing can significantly inhibit the proliferation ability and migration ability of human tongue squamous cell carcinoma cell lines CAL-27.