1.The experimental study of recombinant adenovirus-mediated human wild type p53 gene transfection into neuroblastoma cell line
Journal of Chinese Physician 2008;10(11):1475-1477
Objective To observe the effect of transduetion of human wild type gene(wt-p53) mediated by recombinant adenovirus on cell growth, cell cycle and apoptosis of neuroblastoma (NB) cell line SH-SY5Y. Methods The NB cell line SH-SY5Y were transfected by recombinant adenovirus with green fluorescent protein(Ad-GFP). The suitable muhiplieity of infection (MOI) was determined after trans-fer efficiency was tested by flow cytometry (FCM). Both recombinant adenovirus(Ad) and recombinant adenovirus mediating wt-p53 gene (Ad-p53) were transfected into SH-SY5Y cells. SH-SY5Y cells without any transfection were used as blank group. The p53 protein expres-sion was validated by Western blot. Cell number was counted and growth curve was drawn. Both cell cycle and apoptesis were detected by FCM. Results The results indicated that the transfer efficiency of recombinant adenovirus to SH-SY5Y was satisfied, and 100 MOI was suitable. After transfection with Ad-p53, p53 protein was high expressed. Cell growth were inhibited, apoptosis and cell cycle arrest were in-duced in p53 gene transfeetion group. Conclusions Our results indicate that clinical application of the in vitro p53 gone modified tumor cell vaccine might be feasible and effective in clinical treatment of NB.
2.Effect of Immunogenicity of Neuroblastoma Cell Lines by Transduction of Adenovirus-Mediated Co-Stimulated Factor B7-1 and Granulocyte-Macrophage Colony-Stimulating Factor Genes
Journal of Applied Clinical Pediatrics 1992;0(05):-
ObjectiveTo observe the effect of co-stimulated factor B7-1 and granulocyte-macrophage colony-stimulating factor(GM-CSF) genes mediated by recombinant adenovirus on immunogenicity of neuroblastoma(NB)cell line SH-SY5Y.MethodsAfter NB cell line SH-SY5Y was transfected with recombinant adenovirus mediating green fluorescent protein(Ad-GFP) in different multiplicity of infection(MOI),the transfer efficiency was tested by flow cytometry(FCM).SH-SY5Y cells were transfected separately with recombinant adenovirus,recombinant adenovirus mediating B7-1 and GM-CSF genes or only B7-1 gene,and they were named as Ad,B7-GM and B7 group.The cells which were not transfected were named as SH group.The protein expression of B7-1 and GM-CSF were detected by using FCM and enzyme linked immunosorbent assay(ELISA).Peripheral mononuclear cells were separated and purified with Ficoll density gradient centrifugation.Cpm and cytokine(IFN-? and IL-2) secretory were evaluated by mixed lymphocyte culturing with different ratio(5 to 1 and 10 to 1) on target cells and effect cells.ResultsTransfer efficiency of recombinant adenovirus to SH-SY5Y was high.After transfection,B7-1 and GM-CSF genes were expressed.Proliferation of lymphocytes was stimulated and cytokine increased vigorously in B7-GM group and B7 group,and the effect in B7-GM group was stronger than that in B7 group.And the effect in ratio 5 to 1 group was stronger than that in 10 to 1 group.ConclusionsB7-1 and GM-CSF genes have synergistic effect.After transfection with recombinant adenovirus mediated B7-1 and GM-CSF genes,the immunogenicity of SH-SY5Y is greatly enhanced.
3.Applications and investigations of nano-bone implants
Orthopedic Journal of China 2009;17(21):1632-1635
In recent years,nano - materials have represented the superior biocompatibiliry and the arena of medical nano - materials have been extensively studied by various means all over the world. Is it possible that nano - bone implants overcome the existing limitations of the traditional bone implants for fusing operations in the future? State Food and Drug Administration (SFDA, CN) authorized the nano-bone implants- - nHA/ PA66 (YZB/0063 -2003) in 2005 but there were still lots of counterviews in this field. This review considers the advanced researches via comparing and analyzing the correlatire literatures recently.
4.Effects of wt-p53, BT-1 and CM-CSF genes cotransfection on proliferation and immunogenicity of SH-SY5Y cell lines
Fang CHEN ; Lei LIU ; Bin WANG
International Journal of Pediatrics 2009;36(2):116-119
Objective To observe the effeet of cotransduction of wt-p53,B7-1 and GM-CSF genes mediated by recombinant adenovirus on cell growth, apoptosis and immunogenicity of neuroblastoma cell line SH-SY5Y, Methods We transfected SH-SYSY cells separately with recombinant adenovirus, recombinant adenovirus mediating human wild type p53(wt-p53) gene and BB-102.We separately named them with SH/Ad,SH/p53 and SH/BB-102 group.The protein expression of p53, B7-1 and GM-CSF were measured by Western blot, FCM and enzyme linked immunosorbent assay. Cell number was counted and growth curves were drawed. Cell apoptosis was tested by FCM. Lymphocyte proliferation and cytokine secretory was evaluated by mixed lymphocyte culturing. Results After transection with BB- 102, p53, B7-1, and GM-CSF, high-efficiency expression of target genes were found. Growth of the cells was inhibited, apoptosis was induced in the SH/p53 and SH/BB-102 groups. Proliferation of lymphocytes was stimulated and cytokine increased visorously in the SH/BB-102 group. Condusions After transfection with BB-102, growth of the SH-SY5Y cells was inhibited,apoptosis was induced,and immunogenicity was greatly enhanced.
5.The effects of Streptococcus sobrinus on corrosion behavior of metal-ceramic alloys
Lei CHEN ; Hui CAI ; Changyun FANG
Journal of Practical Stomatology 1996;0(02):-
Objective:To study the corrosion resistance of four metal-ceramic alloys in the presence of Streptococcus sobrinus, and to compare the effects of bacteria action in liquid and solid environment.Methods:Four metal-ceramic alloys were exposured to BHIA or BHI media with or without Streptococcus sobrinus and served as different groups. After inoculating 10 weeks, their I_ corr ,E_ corr ,E_ tp through cyclic polarization curves were obtained through Corrview 2 corrosion analyses software;and their surface changes after electrochemical corrosion were observed by field emission scanning electron microscopy (FESEM).Results:The E_ corr of four alloys were decreased in presence of oral microorganism, and the declines were more remarkable in BHI with bacteria. SEM observation showed the pittings in their surface.The corrosion resistance in BHI without bacteria were significant increased than that in the blank control. Conclusion:Streptococcus sobrinus can promote the corrosion behavior of metal-ceramic alloys and the corrosion resistance is more worse in BHI than BHIA with bacteria.
6.The use of magnifying chromoendoscopy in early detection of gastric cancer and its precancerous lesions
Lei CHEN ; Dianchun FANG ; Xianghong LI
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective To explore the pit patterns of mucosal atrophy, intestinal metaplasia, dysplasia of gastric mucosa and early gastric cancer by magnifying chromoendoscopy. Methods The pit patterns of gastric mucosa of 594 patients with gastrosia were observed with electronic magnifying endoscopy (Olympus GIF Q-240Z) assisted with methylene blue staining. Biopsies were taken from suspicious sites as observed after magnification for histopathologic examination. Results The diagnostic sensitivity, specificity and accuracy of magnifying chromoendoscopy for mucosal atrophy were 74.6%, 90.4% and 84.1%, respectively. In the central part of mild atrophic areas gastric pits were found to be decreased in quantity or even disappeared, while in severe atrophic area, scar-like changes were seen due to the disappearance of pits, with elongation and tortuosity of surrounding. Mucosa with intestinal metaplasia was lightly stained by methylene blue, and pit patterns appeared as villi, scars or coarse lines. Mucosa of dysplasia or early gastric cancer was heavily stained by methylene blue, and gastric pits were amorphous and irregular in size. Conclusion Gastric mucosal atrophy, intestinal metaplasia, dysplasia and early gastric cancer have their peculiar pit patterns, and magnifying chomoendoscopy may be useful in the diagnosis of these lesions.
7.Clinical efficacy of programmed intermittent epidural bolus and continuous epidural infusion for labor analgesia
Xiangdong FANG ; Lei XIE ; Xianxia CHEN
The Journal of Clinical Anesthesiology 2016;32(8):757-760
Objective To compare the clinical efficacy of programmed intermittent epidural bollus (PIEB)+patient controlled epidural analgesia (PCEA)and continuous epidural infusion (CEI)+PCEA for labor analgesia.Methods One hundred nulliparous parturients with cervical dilation of 2-3 cm were randomly assigned to PIEB group or CEI group for labor,with 100 parturients in each group.The background infusion in PIEB group delivered 8 ml bolus at a rate of 6 ml/min per 1 hour, in group CEI at 8 ml/h.Blood pressure,VAS score and modified Bromage score before labor anesthe-sia (T0 ),10 min after labor anesthesia (T1 ),30 min after labor anesthesia (T2 ),1 h after labor an-esthesia (T3 ),2 h after labor anesthesia (T4 ),the uterus opening to the full extent (T5 ),childbirth (T6 ),1 h after childbirth (T7 ),proportion of PCEA request,hourly ropivacaine and sufentanil con-sumption,uterine contraction,fetal heart,total delivery time,analgesic time,delivery mode,number of cases using oxytocin,adverse reactions,neonatal Apgar score,maternal satisfaction score were ob-served.Results The VAS score of PIEB group was significantly lower than that of CEI group at T4-T6 (P <0.01).Compared with the CEI group,protortion of PCEA request,the dosage of drug was de-creased obviously in group PIEB (P <0.05),the maternal satisfaction of PIEB group was significantly higher than that of CEI group (P <0.05).There was no significant difference in the blood pressure, duration and interval time of uterine contraction,fetal heart,total delivery time,analgesic time,deliv-ery mode,number of cases using oxytocin,adverse reactions,neonatal Apgar score and weight among two groups.Conclusion Compared with the CEI+PCEA,the need of PCEA and the dosage of drug was decreased obviously,the VAS score was significantly lower,the maternal satisfaction was signifi-cantly higher and adverse reactions did not increase in the PIEB+PCEA.
8.Advances in anti-tumor research of HDAC inhibitors and combination with PI3K inhibitors
Lei HUANG ; Xiao-guang CHEN ; Fang-fang LAI
Acta Pharmaceutica Sinica 2022;57(12):3557-3563
Histone deacetylase (HDAC) is usually abnormally overexpressed, which mainly leads to the transcriptional repression of tumor suppressor genes. Histone deacetylase inhibitors (HDIs) exert anti-tumor biological effects by regulating nucleosome structure, inhibiting HDAC activity, and controlling the expression of tumor suppressor genes. There are currently 5 drugs on the market, but only for peripheral T-cell lymphoma and cutaneous T-cell lymphoma. In solid tumors, most of the HDAC inhibitors used have failed to achieve effective therapeutic effects. Phosphoinositide 3-kinase (PI3K) is the starting node of the PI3K-AKT-mTOR signaling pathway, which plays a very important role in the proliferation, migration, invasion, and differentiation of tumor cells. The abnormal activation of PI3K is closely related to the occurrence and development of tumors, and the combined use of HDAC and PI3K inhibitors and HDAC/PI3K dual-target inhibitors show synergistic anticancer activity. This article introduces the anti-tumor clinical and preclinical research progress of representative HDAC inhibitors and PI3K inhibitors, as well as HDAC/PI3K dual-target inhibitors.
9.Effect of puerarin on myocardial damage in STZ-induced diabetic rats
Xiufang CHEN ; Kangfu LEI ; Min DONG ; Zhouxi FANG ; Liqin JIN
Chinese Journal of Pathophysiology 2010;26(4):650-655
AIM: To investigate the effect of puerarin on diabetic myocardial damage and to explore its possible mechanisms in rats. METHODS: The male Sprague-Dawley rats were randomly divided into normal control group, diabetic group, high dose puerarin (160 mg/kg), middle dose puerarin (120 mg/kg), low dose puerarin (80 mg/kg) treatment groups and aminoguanidine (100 mg/kg) treatment group. Corresponding drugs were intraperitoneally injected once a day. The animals in normal control group and diabetic group were given equal propylene glycol. 12 weeks later, the rats were sacrificed and their cardiac muscles were collected. Myocardial structure was observed under transmission electron microscope (TEM). The blood glucose concentration and the activities of superoxide dismutase (SOD), Ca~(2+)-ATPase, Na~+-K~+-ATPase and the content of malondialdehyde (MDA) in myocardial homogenate were measured biochemically. In addition, the mRNA expressions of peroxisome proliferator activated receptor γ (PPAR-γ), glucose transporter-4 (GLUT-4) and aldose reductase (AR) in myocardial tissues were determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: It is observed that the myofibrils were diminished, broken or fused, some lipid droplets deposited in the cytoplasm of cardiocytes and part of cristae of mitochondria were broken or disappeared under TEM in diabetic group. The activities of SOD, Ca~(2+)-ATPase, Na~+-K~+-ATPase as well as the mRNA expressions of PPAR-γ and GLUT-4 decreased significantly (all P<0.01). The blood glucose concentration and the MDA level and the AR mRNA expression increased obviously (all P<0.01) in diabetic group as compared to those in normal control group. How-ever, in puerarin treatment groups, the above changes were reversed, a significant differences of those were found as compared to those in diabetic group (P<0.05 or P<0.01, respectively). The pathological change of cardiac muscles was relieved. It showed that myofibrils were well-arranged and only few lipid droplets deposited in the cytoplasm of cardiocytes and most mitochondria had clear and regular cristae under TEM in puerarin treatment groups. CONCLUSION: Puerarin exerts preventive and remedial effects on the diabetic myocardium, which may be related to up-regulating the mRNA expressions of PPAR-γ and GLUT-4, promoting glucose uptake and relieving oxidative stress damage.
10.Investigation on epidemic status of clonorchiasis sinensis based on inpatients in Zhongshan City,China
Man WANG ; Le LUO ; Xueqin CHEN ; Lei LI ; Yueyi FANG
Chinese Journal of Schistosomiasis Control 2017;29(4):459-463
Objective To understand the prevalence and epidemiological features of clonorchiasis sinensis based on inpa-tients in Zhongshan City. Methods All of the 23 town hospitals in Zhongshan City were selected as the surveillance hospitals in 2016. Among the inpatients from those hospitals,those lived in the city longer than 6 months were selected as the surveillance subjects. The stool specimens of the subjects were collected and examined by the modified Kato-Katz thick smear method for the eggs of Clonorchis sinensis. The crude infection rates of C. sinensis of the subjects were standardized according to the age and sex compositions of the population in Zhongshan City,2016. Meanwhile,the infection rates of the subjects from different towns and those combined with related clinical diseases were analyzed. Results A total of 2667 people were included in the surveil-lance,among which 219 were tested as positives,with a crude infection rate of 8.21%. For the positives,the male accounted for 81.74%(179/219),and their age ranged from 4 to 89 years with the mean value of(62.20±13.80)years. After standardization by age and sex composition,the infection rate was 3.62%for the whole population,the rates of the male and the female were 5.46%and 1.53%,respectively,and the former was 3.57 times as high as the latter. Both the infection rates of the male and the female rose as the age increasing(χ2 male=99.91,χ2 female=16.51,both P<0.01). The standardized infection rates among the subjects in different towns ranged from 0 to 10.40%,and the rates in the north towns were higher than those in the south towns. The standardized rate was 9.22%for the subjects combined with gallstone,3.40%for those not combined with gallstone,and the former was 2.71 times as high as that of the latter. Conclusion The people aged 30 years and above,with gallstone or living in towns with high C. sinensis infection rates are the key population for prevention and control of clonorchiasis in Zhongshan City.