1.Construction of lentiviral vector pLeno-DCE-PDGF-B and its effect on invasion ability of SGC7 9 0 1 gastric carcinoma cells
Chongqing Medicine 2014;(18):2260-2263
Objective To construct a lentiviral vector that stably express platelet-derived growth factor B (PDGF-B)and to in-vestigate effect of PDGF-B overexpression on invasion of SGC7901 gastric carcinoma cell.Methods The lentiviral vector pLeno-DCE-PDGF-B was constructed and transfected into 293T cells.The supernatant containing the lentivirus particles was harvested to determine the virus titer and high titer lentivirus particles was gathered.Then PDGF-B lentiviral vector was transfected into SGC7901 cells for construction of stable PDGF-B overexpression SGC7901 cells.Western-blot and immunofluorescence were used for evaluation of the construction of stable PDGF-B overexpression SGC7901 cells.Transwell test was used for detection the inva-sion of SGC7901 cells.Results The lentiviral vector was correctly constructed and verified by sequencing.High titer PDGF-B lenti-viral particles were acquired successfully.After transfection,SGC7901 cells were green color by detection of fluorescence micro-scope;and the expression of PDGF-B protein in transfected SGC7901 cells were much higher than that in normal SGC7901 cells.Al-so,PDGF-B overexpression increased the invasion ability of SGC7901 cells.Conclusion The PDGF-B lentiviral particles are ob-tained after successful construction and package.The stable PDGF-B overexpression SGC7901 gastric carcinoma cell line is estab-lished,which proves that the PDGF-B overexpression might increase the invasion ability of gastric carcinoma cells.
2.Investigation in depression emotion, personality and quality of life of patients with digestive system tumor undergoing chemotherapy
Miao ZHANG ; Weili WANG ; Jiangyan SONG
Chinese Journal of Practical Nursing 2011;27(25):1-5
ObjectiveTo study the state of depression emotion,personality and quality of life of patients with digestive system tumor undergoing chemotherapy, and the correlation of depression emotion with personality and quality of life.MethodsUsing cross-sectional survey methodology, 261 digestive system tumor patients were investigated by schedule table SDS, EPQ-RSC and EORTC QLQ-C30. These cases were also used to study the correlation of depression emotion with personality and quality of life.ResultsThe digestive system tumor patients undergoing chemotherapy investigated had a high score in depression than domestic norm, and the incidence of depression of these cases was 19.5%.The cases were divided into two groups(the depression group and the non-depression group). There were significant differences in N and P dimensionalities of personality and in physical function, role function, cognitive function, emotional function,and overall well-being, fatigue, economic difficulties, nausea and vomiting, pain, insomnia, loss of appetite,constipation and diarrhea. There were various degrees of difference in depression, personality and quality of life.ConclusionsDepression, personality and quality of life are closely related in digestive system tumor patients undergoing chemotherapy. The medical staff should pay more attention to the patients quality of life and personality to make a better nursing intervention and improve their quality of life.
3.Efficacy of 32P and pinyangmycin injection in combination on maxillofacial hemangioma in infants and children
Jingui HU ; Xuehong CHEN ; Jiangyan LIU ; Zhengjiang WANG ; Xiujuan LU
Chinese Journal of Medical Aesthetics and Cosmetology 2010;16(5):323-325
Objective To explore and evaluate the effect of 32P and pinyangmycin injected in combination for the treatment of maxillofacial hemangioma. Methods The patients were randomly divided into three groups. The first group of patients was treated with 32P; 100 patients were enrolled in this group, of which 37 were male and 63 female. 32P dosage between 0.37 and 0. 74 MBq per cm3 was prepared together with 0.5 ml of 2 %lidocaine, and appropriate normal saline, and then the mixture was injected into hemangioma every two weeks. The second group was given pinyangmycin, and the third group received both 32P and pinyangmycin. The second group included 30 male and 60 female, and the third group 64 male and 136 female. The dose and frequency were given as the same to the first group. Results 88.5 % of patients were cured using both 32P and pinyangmycin, and the cure rate was 77.0 % and 71.7 % with either 32P or pinyangmycin. Conclusion These results prove that 32P and pinyangmycin injected in combination for the treatment of maxillofacial hemangioma is more effective than either 32P or pinyangmycin alone, and furthermore, the method is easy, safe and has less complications.
4.A novel 5' splice site mutation in ACTC1 gene may play an important role in congenital ventricular septal defect
Hang LI ; Binbin WANG ; Bingren GAO ; Jiangyan LIU ; Qiming ZHAO ; Wensheng CHEN ; Wei WANG ; Kun YANG
Chinese Journal of Tissue Engineering Research 2012;16(11):2087-2090
BACKGROUND: As a candidate gene of congenital heart disease, ACTC1 gene is related to congenital atrial septal defect inhumans.OBJECTIVE: To perform a mutation screen of ACTC1 gene in 110 nuclear families of congenital heart disease.METHODS: A case-control study was conducted based on 110 nuclear families of congenital heart disease and 300 normalhuman beings with no reported cardiac malformation. Six fragments in the coding region of ACTC1 gene was amplified by PCR invitro using five primers pairs. PCR products were screened for gene mutations.RESULTS AND CONCLUSION: A novel G-to-A variant was found at the third nucleotide of the intron downstream from exon 5.This mutation existed in a 5-year-old female with an isolated ventricular septal defect and her 30-year-old father, who had noreported cardiac anomalies. This mutation was not detected in 300 normal controls. These findings indicate that the mutation maybe related with congenital ventricular septal defects in humans.
5.Multiple insecticide resistances and kdr mutation of Culex pipiens pallens in north-central Anhui Province
Xing WEI ; Xiaodi YANG ; Xiaoli WANG ; Jiangyan LI ; Yuanyuan WANG ; Hui XIA ; Xuelian CHANG ; Qiang FANG
Chinese Journal of Schistosomiasis Control 2016;28(4):406-410
Objective To understand the status of resistance to multiple insecticides and the frequencies of kdr mutations in Culex pipiens pallens from north?central Anhui Province. Methods From July to September,2014,the C. pipiens pallens mosquito larvae were collected in Huaibei,Bengbu and Chuzhou cities of the north?central Anhui Province and reared to adults. The female adult mosquitoes at 3-5 days post emergence were tested for susceptibility to the four insecticides,namely 0.05%deltamethrin,5%malathion,0.1%bendiocarb and 4%DDT,by using the standard WHO resistance tube bioassay. The detec?tion of the point mutations of the kdr gene at codon 1014 was conducted by PCR and DNA sequencing in the deltamethrin?resis?tant and?susceptible mosquitoes. Results High levels of resistance to all the four insecticides were found in all the three tested populations,although mosquito mortality varied among populations and test insecticides. Among the test insecticides,DDT showed lowest mortality with no significant difference(F=1.027,P>0.05)in all test populations,whereas significantly differ?ent mortalities were observed among populations for the remained three insecticides tested(deltamethrin,malathion,and ben?diocarb)(F = 23.823,33.955,128.841;all P < 0.01). Two types of non?synonymous kdr mutation at codon position 1014 (L1014F and L1014S)were observed. A positive correlation between L1014F mutation frequencies and deltamethrin resistance levels were detected in the three mosquito populations(r2=0.718,P<0.01). Conclusions The observed high levels of resis?tance to multiple?insecticides coupled with the occurrence of medium to high kdr frequencies in populations of C. pipiens pallens could profoundly affect the mosquito vector control programme in China. The local health departments need to strengthen vector dynamic monitoring and implement rational resistance management strategies.
6.Dynamic monitoring of donor specific antibodies in living-relative renal transplantation and early intervention
Yu CHEN ; Jiangyan LI ; Xiaoyan LOU ; Hao WANG ; Lei ZHANG ; Youhua ZHU
Chinese Journal of Organ Transplantation 2014;35(5):267-269
Objective To analyze the clinical application of HLA donor specific antibodies (DSAs) detected by Luminex single antigen beads,and to discuss the impact of early intervention on renal function.Method In 64 cases of living-relative renal transplantation,DSA was detected using a Luminex single antigen assay before and after transplantation.The positive recipients were given large doses of intravenous irnmunoglobulin (IVIG) and increased doses of mycophenolate mofetil (MMF).The relationship between DSA and renal function was analyzed.Result DSA was negative in all recipients before transplantation.Ten cases of DSA positive recipients were found in HLA mismatch after transplantation.After the intervention,two cases of DSA positive recipients became negative,immunofluorescence intensity was decreased by more than 50% in 6 cases,and no significant reduction was found in the other two cases.Antibody-mediated rejection (AMR) occurred in two cases of intervention ineffective recipients after 3 to 6 months and the renal function was impaired.Conclusion Dynamic monitoring of DSA using Luminex single antigen beads may timely predict changes of renal function.Early application of large doses of IVIG and increasing doses of MMF can reduce the incidence of AMR.
7.Construction and identification of recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 antigen
Xuemei WANG ; Jiangkun LUO ; Qian LI ; Jiangyan LI ; Yong CHEN ; Zhiyong TAO ; Hui XIA ; Qiang FANG
Chinese Journal of Schistosomiasis Control 2014;(3):287-291
Objective To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 anti-gen. Methods The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction en-zymes,and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both frag-ments were modified by Klenow fragment to form blunt end,then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1-Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction,a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band,which sug-gested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis,the recombi-nant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombi-nant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.
8.Long-term Effects of Neonatal Seizures and Exercise on Learning,Memory and CaMKⅡ Expression in Hippocampus
Hong NI ; Yuwu JIANG ; Luyang TAO ; Jiangyan LOU ; Zhedong WANG ; Xiru WU
Progress in Biochemistry and Biophysics 2006;0(01):-
Despite the clinical and experimental concerns about the deleterious effects of neonatal seizures on brain development,the underlying mechanism of seizure-induced brain damage is still not clear.Moreover,early therapeutic intervention studies are also less available.For this reason,the study was performed to explore the long-term effects of neonatal seizures and physical exercise on learning,memory and the expression of calcium/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ).Twelve neonatal rats for each group were assigned:the single-seizure group(SS),the recurrent-seizure group(RS) and the control group.The volatile agent flurothyl was used to induce 30 min seizure attack.At postnatal day 6(P6),the single seizures induced only once and recurrent seizures induced once per day for consecutive 6 days.Control rats were placed into the container for an equal amount of time to their counterpart without exposure to flurothyl.Morris water-maze test were performed at P27~P31,P58~P61 and P80~P82,meanwhile at P51~P56,the RS and SS groups were submitted to forced running exercise.In situ hybridization method was used to detect the expression of CaMKⅡ mRNA in hippocampus.The results are as follows:(1) Escape latency.In the first two Morris water-maze tests,there was a decreasing trend of escape latency in three groups,and the escape latency of RS group was much longer than that of control group.After physical exercise,in the last Morris water-maze tests,the diference of escape latency in three groups is not significant.(2) Searching strategy.In the first Morris water-maze test,there was a decreasing trend of marginal strategy and an increasing trend of taxis strategy in three groups,but the frequency of marginal strategy was higher and the frequency of taxis strategy was lower in RS group than that in SS and control group in the third and fourth day(P
9.Regulation and its signaling mechanism of miR-328on endothelial-mesenchymal transition induced by high glucose of humanumbilical vein endothelial cells
Liang WANG ; Yunxiao CHEN ; Junfang WU ; Hong JIANG ; Huan WAN ; Jiangyan HUANG ; Jun HUANG
Basic & Clinical Medicine 2017;37(8):1098-1102
Objective The study is to investigate the role of miR-328 in endothelial mesenchymal transition (EMT)induced by high glucose in human umbilical vein endothelial cells (HUVECs)and its signaling mechanism.Methods HUVECs were cultured in high glucose environment to induce EMT;The recombinant lentiviruses were created by miR-328 and antagomiR- 328 transfection of HUVECs.The experiment was divided into seven groups: normal glucose;mannitol group;high glucose;miR-328;miR-328 virus negative control;high glucose + U0126;miR-328 + U0126.Double immunofluorescent staining was used to determine expression of EMT markers;Changes in miR-328 expression is examined by RT-qPCR;The expressions of type Ⅰ/Ⅲ collagen,p-MEK1/2 and p-ERK1/2 are examined by Western blot.Results 1)HUVECs showed positive staining for CD31 and α-SMA in high glucose group.2)The expression of miR-328 was up-regulated(P<0.05)in HUVECs treated by high glucose or miR-328.Compared with high glucose group or miR-328 group,miR-328 expression was less pronounced aftertreatment with U0126.3)The expressions of type Ⅰ/Ⅲ collagen increased in HUVECs treated by high glucose or miR-328(P<0.05) Compared with high glucose group or miR-328 group,typeⅠ/Ⅲ collagen expressions were less pronounced after treatment with U0126.4)The expressions of p-MEK1/2 and p-ERK1/2 were increased in HUVECs treated by high glucose or miR-328 in comparison to the control group (P<0.05);a lower expression of p-MEK1/2 and p-ERK1/2 were observed in U0126 group than in high glucose group or miR-328 group.Conclusions The phenomenon of EMT in HUVECs is induced by high glucose with increased expression of miR-328;overexpression of miR-328 induced EMT in HUVECs;miR-328 induced EMT is related with MEK1/2-ERK1/2 signaling pathway.
10.SPECT and CT in Evaluating Preoperative Renal Function of Patients with Unilateral Hydronephrosis
Xiaoxi PANG ; Hao ZHONG ; Xuehong CHEN ; Zhengjiang WANG ; Jiangyan LIU ; Junlin ZHOU
Chinese Journal of Medical Imaging 2015;(2):120-124
Purpose To analyze the relationships between glomerular filtration rate (GFR) measured by SPECT and renal parenchyma thickness and enhanced CT value measured by CT, and to explore predictive significance of CT in evaluating renal function of patients with hydronephrosis.Materials and Methods One hundred and fifteen patients diagnosed with unilateral hydronephrosis by ultrasound were retrospectively analyzed. GFR% (GFR percentage of affected kidney to the both kidneys) was measured by SPECT. CT% (percentage of affected renal parenchyma thickness to the both kidneys) and enhanced CT% (percentage of enhanced CT value of affected kidney to the both kidneys) were measured by pre- and post-contrast CT scan. According to GFR, the renal function was divided into mild-to-moderate impairment, severely impairment and non-function. Twenty-five volunteers were recruited as control group. CT%, enhanced CT% and GFR%among the four groups were compared, and the correlation of CT% and enhanced CT%with GFR% was analyzed to evaluated CT in predicting renal function.Results CT%, enhanced CT% and GFR% in mild-to-moderate impairment group was significantly greater than those in severely impairment group and non function group (F=20.24, 7.78 and 329.21,P<0.05). GFR% was positive correlated with CT% (r=0.58,P<0.05) and enhanced CT% (r=0.61,P<0.05). Area under curve (AUC) of CT% were 0.54, 0.79 and 0.83 for mild-to-moderate impairment, severely impairment and non-function, with sensitivity of 92.91%, 93.47%, 65.72%, and specificity of 35.33%, 59.47%, and 88.62%, respectively. AUC of enhanced CT% were 0.79, 0.89 and 0.96 for the three groups, with sensitivity of 97.51%, 80.02%, 97.66%, and specificity of 58.14%, 89.82% and 94.27%, respectively.Conclusion There was high correlation between renal function imaging by SPECT and CT in evaluating renal function of hydroneohrosis patients. Pre- and post-contrast CT scan can be used as complements in predicting renal function, and post-contrast CT with high accuracy.