1.Screening of marker gene in Yersinia pestis plasmid
Shan-shan, DONG ; Ying, GUO ; Peng, WANG ; Zhi-zhong, SONG
Chinese Journal of Endemiology 2012;31(5):490-494
Objective To screen the conservative,stable and specific DNA signature sequence in the plasmid of Yersinia pestis.Methods Specific validation trials and stability of the qualification test were carried out to 40 strains of Yersinia pestis,47 strains of non-Yersinia pestis of home and wild types of rodent in Yunnan,by using 32 DNA sequences derived from Yersinia pestis in the plasmid and conventional PCR technology,and Yersinia pestis vaccine strain EV76 as a positive control.Results Four pairs of relatively conservative,stable and specific DNA marker genes were screened:YPMT1.05c,YPMT1.03c,YPMT1.42 and YPMT1.04c.Conclusions The 4 pairs of Yersinia pestis DNA signature sequences can be used for rapid diagnosis of plague.
2.Co-treatment of endothelial progenitor cells and pioglitazone improves kidney function in diabetic rats
Song WANG ; Xisheng ZHENG ; Shan WANG ; Fang FANG
Chinese Journal of Tissue Engineering Research 2017;21(17):2702-2707
BACKGROUND: Pioglitazone is a common hypoglycemic drug capable of improving proliferation and activation, and inhibiting apoptosis of endothelial progenitor cells (EPCs). We speculated that the combined use of pioglitazone and EPCs transplantation could have significant improving effects on hyperglycemia and kidney function after diabetes mellitus.OBJECTIVE: To investigate the improving effect of EPCs transplantation combined with pioglitazone treatment on the kidney function of diabetic rat models.METHODS: The 15 of 75 Wistar rats were randomly selected and served as normal control group (no treatment). Animal models of type 1 diabetes mellitus were made in the rest 60 rats through the intraperitoneal injection of 40 mg/kg streptozotocin for continuous 5 days. The human EPCs (labeled by CM-Dil) were recovered, cultured and preserved until transplantation. After 4 weeks of modeling, the 60 rat models were randomly divided into model group (PBS injection), pioglitazone group, EPCs transplantation group and combined treatment group, followed by tail vein injection of EPCs suspension and/or intragastric administration of 20 mg/kg pioglitazone for continuous 4 days. After 8 weeks of treatment, the levels of glucose, insulin and creatinine in serum, urea nitrogen and urine protein during 24 hours were determined. The number and distribution of EPCs labeled by CM-Dil were detected by fluorescence microscope, the apoptosis in kidney cells was tested by TUNEL method, and the kidney weight/body weight ratio in rats was calculated.RESULTS AND CONCLUSION: Compared with the model group, the blood glucose and serum creatinine levels and the urea nitrogen and urine protein concentrations during 24 hours were significantly decreased (P < 0.05), and the seruminsulin level was significantly increased (P < 0.05) in the pioglitazone and EPCs transplantation groups. These biochemical indexes in the combined treatment group were more significantly altered compared with the model group (P< 0.01). The kidney weight to the body weight ratio was lowest in the combined treatment group and lower in the pioglitazone and EPCs transplantation groups followed by the model group (P < 0.05). The order of apoptotic kidney cells labeled by TUNEL was as follows: model group > pioglitazone group > EPCs transplantation group > combined treatment group (P < 0.05). To conclude, the EPCs transplantation combined with pioglitazone treatment can decrease the blood glucose and serum creatinine levels and urea nitrogen and urine protein concentrations, improve the serum insulin level, reduce cell apoptosis in the kidney, and remit the kidney dysfunction of diabetic rats to a certain extent.
4.A case report of lipomatous hemangiopericytoma
Liru DONG ; Huiqiang REN ; Shan WANG ; Xudong SONG
Tianjin Medical Journal 2016;44(10):1293-1294
Lipoma of hemangiopericytomas (LHPC) is one ofrare soft-tissue tumors that grow slowly and occur in deep soft tissues. The classical histological morphology of LHPC is circularor spindle undifferentiated tumor cells that grow surrounding the thin-walled branching blood vessels. The clinical and pathological features of one patient with LHPC are retrospective analyzed to strengthen the understanding of LHPC.
5.Effects of phenytoin on VEGF and SCF in rat bone marrow mesenchymal stem cells and vascular endothelial cells co-culture system
Shan HE ; Liyue ZHAO ; Xiaoye BA ; Baoyan WANG ; Jianling SONG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;(2):204-208,225
ABSTRACT:Objective To investigate the effects of phenytoin (PHT)on the secretion of vascular endothelial growth factor (VEGF)and stem cell factor (SCF)based on the establishment of indirect co-culture system of rat bone marrow mesenchymal stem cells (BMSCs)and vascular endothelial cells (VECs).Methods Indirect co-culture model of rat BMSCs and VECs was established.Experimental groups:indirect co-culture groups (PHT concentrations were 0,20 and 40 μg/mL);the control group:BMSCs culture group and VECs culture group (PHT concentrations were 0,20 and 40μg/mL).The contents of VEGF and SCF in the culture supernatant were measured using double antibody sandwich ABC-ELISA method on cultivation days 2,4,6.Results ELISA assay of the rBMSCs and rVECs in indirect co-culture supernatants,collected on culture days 2,4 and 6 showed that:① VEGF:On culture day 2,VEGF level in the co-culture groups was significantly higher than those in BMSCs group (P <0.05)and rVECs group (P <0.001).As culture time prolonged and PHT concentration increased to 20 μg/mL and 40 μg/mL,VEGF level increased too (P <0.001,P <0.05 ).② SCF testing results showed that the secretion of SCF in co-culture groups was higher than that in the control groups.When PHT was 20 μg/mL,the secretion of
SCF increased as the incubation time increased;but as the incubation time increased, PHT concentration of 40 μg/mL made SCF content decrease.Each group did not significantly differ (P > 0.05 ).Conclusion PHT promotes the secretion of VEGF and may reduce the secretion of SCF.
6.Cardiac repair effect of AKT transfected amniotic fluid mesenchymal stem cells on ischemic reperfusion ;injured myocardium in rabbits
Lei SONG ; Song ZHANG ; Yan WANG ; Shan JIANG ; Yanyan LI ; Wei LIU ; Yingmin LU
Chinese Journal of cardiovascular Rehabilitation Medicine 2017;26(1):68-73
Objective:To explore cardiac repair effect of AKT gene transfected amniotic fluid mesenchymal stem cells (AFMSC) transplantation on ischemic reperfusion injured myocardium in rabbit model . Methods :AKT overex‐pressed lentiviral vector was established to transfect AFMSC ;New Zealand rabbits were randomly divided into low glucose DMEM (L‐DMEM) group (group A) ,AFMSC group (group B) and AKT transfected AFMSC group (AKT‐AFMSC group ,group C) .Then rabbit myocardial ischemia reperfusion injury model was established .Before reper‐fusion ,0.2ml L‐DMEM ,AFMSC and AKT‐AFMSC was directly injected into infarct area and surrounding myocar‐dial epicardium in corresponding group . On 21 days after operation , left ventricular end‐diastolic dimension (LVEDd) ,left ventricular shortening fraction (LVFS) ,left ventricular ejection fraction (LVEF) ,infarct size and pathological changes were compared among three groups .Results:Compared with L‐DMEM group and AF‐MSC group ,there was significant reductions in LVEDd [ (16.37 ± 0.84) mm ,(15.06 ± 1.01) mm vs .(13.51 ± 0.85) mm] and infarct size [ (37.3 ± 2.1)% ,(26.6 ± 0.7)% vs .(18.1 ± 1.2)% ] ,and significant rise in LVFS [ (29.18 ± 2.36)% ,(33.65 ± 2.81)% vs .(36.89 ± 3.02)% ] and LVEF [ (58.62 ± 3.47)% ,(67.42 ± 3.03)% vs .(72.02 ± 2.89)% ] , P< 0.05 or < 0.01. Pathological injury significantly relieved in AKT‐AFMSC group than those of group A and group B .Conclusion:Compared with amniotic fluid mesenchymal stem cells ,AKT‐transfected amniotic fluid mesenchymal stem cells possesses better effects of repairing injured myocardium and improving myocardial function in ischemic reperfusion injured myocardium .
7.Effects of sevoflurane preconditioning on postoperative cognitive dysfunction in aged rats
Lili YU ; Aihua ZHAO ; Shuping HUO ; Xuguang CHEN ; Shan SONG ; Qiujun WANG ; Luqi WANG
Chinese Journal of Anesthesiology 2013;33(11):1310-1314
Objective To evaluate the effects of sevoflurane preconditioning on postoperative cognitive dysfunction in aged rats.Methods One hundred and twenty healthy male Sprague-Dawley rats,aged 18 months,weighing 400-450 g,were randomly divided into 3 groups (n =40 each) using a random number table:control group (group C),operation group (group O),and sevoflurane preconditioning group (group Sev).In group C,the rats were only anesthetized with pentobarbital sodium and did not undergo operation.In group O,the rats were anesthetized with pentobarbital sodium and underwent 30 min of exploratory laparotomy.In group Sev,the rats inhaled 2.4% sevoflurane for 30 min and then inhaled air for 30 min,and the other procedures were similar to those previously described in group O.At 30 min before operation and on 1st,3rd,5th and 7th days after operation,Morris water maze test was performed to record the escape latency,time of staying at the original platform quadrant and frequency of crossing the original platform.At 30 min before operation and on 1st and 7th days after operation,10 rats in each group were sacrificed and hippocampi were isolated to detect the apoptotic rate and intracellular [Ca2 +] i (using flow cytometry) and the ultrastructure of hippocampal neurons was observed with transmission electron microscope.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,and the apoptotic rate and intracellular [Ca2 +] i were increased after operation in O and Sev groups (P <0.05).Compared with group O,the escape latency was significantly shorten,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased,and the apoptotic rate and intracellular [Ca2+]i were decreased after operation in group Sev (P < 0.05).Microscopic examination showed no abnormality in the ultrastructure of hippocampal neurons in group C,and the pathological changes of the ultrastructure of hippocampal neurons were obvious in group O,and were significantly attenuated in group Sev.Conclusion 2.4% sevoflurane preconditioning can reduce postoperative cognitive dysfunction in aged rats,and regulation of imbalance of calcium homeostasis and reduction of cell apoptosis are involved in the mechanism.
8.A study of protein expression of MAGE-A3, MAGE-A4 and MAGE-A10 genes in colorectal carcinoma and its clinical significance
Fangfang LIU ; Danhua SHEN ; Shan WANG ; Yingfiang YE ; Hui QIU ; Chenggang WANG ; Yanbin ZHANG ; Qiujing SONG
Chinese Journal of General Surgery 2012;27(1):37-39
Objective To explore protein expression and significance of MAGE genes in colorectal carcinoma(CRC)tissues.Methods The expression of MAGE genes were studied by using tissue chip and immunochemistry methods in primary CRC tissue and paired adjacent tissue samples in 97 cases.Data were analyzed with x2-test by SPSS 16.0 software.Results The protein expression of MAGE-A3,MAGE-A4 and MAGE-A10 genes were 57%(56/97),63%(61/97)and 28%(27/97)respectively in 97 cases of primary adenocarcinoma.The protein expression frequency of MAGE-A3 in poor colorectal adenocarcinomas was significantly higher than in well-and moderately disfferentiated adenocarcinomas(x2 =9.133,P =0.010).MAGE-A10 in poor colorectal primary adenocarcinomas was significantly higher than in well and moderately adenocarcinomas(x2 =15.280,P =0.000); MAGE-A10 protein expression was significantly higher in stage TNM Ⅲ + Ⅳ than in stage TNM Ⅰ + Ⅱ(x2 =4.227,P=0.040); MAGE-A10 gene expression was higher in metastasis lymphoid node than in no metastasis lymphoid node(x2 =5.557,P =0.018),and the expression level was higher in primary lesion with the increasing of the numbers of lymphoid node metastasis(x2 =7.296,P =0.026).Conclusions The protein expression of MAGE genes is associated with the tumor differentiation,TNM stage and lymphoid node metastasis.MAGE-A3 and MAGE-A10 genes are the possible prognosis marker and potential target of immunotherapy of CRC.
9.The protein expression of PLAC1/CP1 genes in colorectal carcinoma and its clinical significance
Fangfang LIU ; Danhua SHEN ; Shan WANG ; Yingjiang YE ; Chenggang WANG ; Yanbin ZHANG ; Qiujing SONG
Chinese Journal of General Surgery 2010;25(12):985-987
Objective To explore the protein expression of PLAC1/CP1 ( cancer-placenta 1 ) and its correlation with clinicopathological characteristics in patients of colorectal carcinoma. Methods The expression of PLAC1/CP1 gene was studied by using tissue chip and immunochemistry in 125 cases CRC tissue specimens. Data were analyzed with the x2-test or Fisher's x2 test statistic by SPSS 16. 0 software.Results The protein expression of PLAC1/CP1 gene was 57.6% (72/125) in 125 cases of CRC and 56. 7% (55/97)in 97 primary adenocarcinoma cases. That was 78. 9% (15/19)in poor differentiated colorectal primary adenocarcinoma, those were significantly higher than that of 35.3% (6/17) in well and 55.7% (34/61) in moderately differentiated adenocarcinoma ( P < 0.05 ); PLAC1/CP1 protein expression was significantly higher in stage TNM Ⅲ + Ⅳ 71.2% (37/52) than in stage TNM Ⅰ + Ⅱ 40% (18/45)(P < 0.05 ); PLAC1/CP1 genes expression rate was 69.6% (32/46) in these with lymphoid node metastasis and 45.1% (23/51)in patients without lymphoid node metastasis (P < 0.05 ). The positive expression rate of PLAC1/CP1 increased in colorectal carcinoma with the increasing of the numbers of lymphoid node involved with metastasis( x2 = 13. 353, P = 0.001 ). Conclusions The protein expression of PLAC1/CP1 is associated with tumor differentiation, TNM stage and lymphoid node metastasis.
10.Clinical Study on Compound Sophora Flavescentis Radix (Kushen) Injection Combined with Gamma Knife for Treatment of Locally Advanced Pancreatic Carcinoma
Lei WANG ; Lingzhao LI ; Xiaofang WANG ; Guoyong SHAN ; Rui LIU ; Song ZHANG ; Xiaodong WU
Chinese Journal of Information on Traditional Chinese Medicine 2015;(6):21-24
Objective To evaluate the clinical efficacy of Compound Sophorae Flavescentis Radix (Kushen) Injection combined with gamma knife for treatment of locally advanced pancreatic carcinoma. Methods Totally 65 patients of locally advanced pancreatic cancer were randomly divided into two groups:32 cases of simple gamma knife group (group A), 33 cases of Compound Kushen Injection combined with gamma knife treatment (group B). Patients received enhancement computed tomography scanning and CA199 review at 1, 3, 6, 9, 12 months after treatment, and once every six months, a total of 36 months of follow-up. Short-term and long-term efficacy was observed after treatment, and survival curve analysis was carried out. Results Clinical benefit response was 53.13% in group A and 84.85% in group B (P=0.006);median survival time of two groups was 11.00 months in group A and 23.00 months in group B;the average survival time in group A was 16.37 months, and 22.28 months in group B (P=0.017). In the two groups, treatment of the 65 patients was completed smoothly as planned, and no one had serious complications, such as pancreatic leakage, skin burns, viscera perforation and radioactive enteritis. Conclusion Compound Kushen Injection combined with gamma knife can treat locally advanced pancreatic carcinoma safely and efficiently, which can improve patients’ life quality, especially in cancer pain control.