1. Construction of lentiviral vector for RNA interference of Id2 gene and its effect on proliferation and apoptosis of gliomas cell line in vitro
Academic Journal of Second Military Medical University 2011;32(6):598-602
Objective To construct a lentiviral vector carrying short hairpin RNA (shRNA) of human Id2 gene and to investigate its effect on the proliferation and apoptosis in glioma cell line U251. Methods Four shRNA sequences targeting human Id2 gene were designed and synthesized. The shRNAs were inserted into the pGCSIL-GFP lentiviral vector. Id2 sequence was amplified from the plasmid which contained Id2 gene as verified by PCR, and was inserted into the pEGFP-N1-3FLAG vector. Lentiviral vector for RNAi of Id2 was constructed and was used to infect 293T cells. The infection efficiency was evaluated and then the lentiviral vector was used to infect U251 cellline. Western blotting analysis was employed to assess the gene silencing efficiency. Cell proliferation was tested by MTT. Cell apoptosis was observed by RT-PCR analysis of caspase 3 expression in U251 cells after infection. Results The results of PCR analysis and DNA sequencing agreed to each other. Compared with the control group, the transfected U251 cells had significantly decreased cell proliferation, and increased apoptosis rate and caspase 3 expression (P<0.05).Conclusion We have successfully constructed lentivirus RNAi vector of Id2, which can inhibit the proliferation and promote the apoptosis of glioma cells in vitro.
2.Change of Urinary Leukotriene E_4 Level in Children with Respiratory Syncytial Virus Bronchiolitis and Its Clinical Significance
Ai-zhen, WANG ; Zhen-yu, ZHANG ; Luo-zhong, ZHAO
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To explore the clinical significance of urinary leukotriene E4(LTE4) in children with respiratory syncytial virus(RSV) bronchiolitis.Methods Forty infants with bronchiolitis were enrolled for the identification of RSV-Ag from the nasopharyngeal secretions.Forty cases who were found to be positive of RSV were put into RSV bronchiolitis group.According to the severity of illness,the infants in RSV bronchiolitis group were separated into 2 subgroups:mild illness group (24 cases) and moderate illness group (16 cases).Thirty healthy children were chosen as healthy control group.Urine of the patients in RSV bronchiolitis group was collected in acute phase (on the 1st day) and recovery phase (on the 7th day after the admission).Urine of children in control group was collected .Enzyme-linked immunosorbent assay was used to assay the level of urinary LTE4.Results The levels of LTE4 in healthy control group (M,Q) were 1.28 pmol?mmol-1,1.72 pmol?mmol-1,while the levels of LTE4 in acute phase in RSV bronchiolitis group (M,Q were 30.52 pmol?mmol-1,23.56 pmol?mmol-1,and the levels of LTE4 in recovery phase in RSV bronchiolitis group (M,Q) were 3.98 pmol?mmol-1,3.43 pmol?mmol-1.Urinary LTE4 levels in acute phase in RSV bronchiolitis group were significantly higher than those in healthy control group (P
3.Effects of ginseng stem and leave saponin on prolactin and menstrual cycle of experimental hyperprolactinemia rats.
Zhen ZHAO ; Yu CAO ; Shu-de YUAN
Chinese Journal of Applied Physiology 2005;21(2):144-195
Animals
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Diestrus
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drug effects
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Female
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Hyperprolactinemia
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physiopathology
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Panax
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Prolactin
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blood
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Rats
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Rats, Wistar
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Saponins
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pharmacology
4.Effects of HSP70 on hepatic ischemia-reperfusion injury after heat preconditioning in rats
Zhen ZHAO ; Dongyan AN ; Lijuan QIN ; Yu CAO
Chinese Pharmacological Bulletin 1986;0(04):-
Aim To study the effects of HSP70 on hepatic ischemia-reperfusion injury after heat preconditioning in rats.Method To establish the models of the hepatic ischemia-reperfusion injury. 42 Wistar rats were randomly divided into 6 groups, (each group had 7 rats): normal group(N); quercetin injection group(Q); ischemia-reperfusion group(I); heat preconditioning 16 hours before ischemia-reperfusion group(H+I); quercetin injection before heat preconditioning group(Q+H+I); quercetin injection before ischemia-reperfusion group(Q+I).We detected the activity of serum enzyme of ALT,AST and the pathological changes of the liver;The expressions of HSP70 of the rats were observed by Western blotting. Results The expressions of HSP70 from high to low were:group H+I,group I,group Q+H+I,group Q+I,group Q,group N; The serum levels of ALT and AST from high to low were: group Q+I,group I,group Q+H+I,group H+I,group Q,group N;All groups had visibly hepatic histological changes respectively.Conclusion The protection of heat stress pretreatment from ischemia reperfusion injury was possibly performed by inducing the expression of HSP70.
5.Dead Reasons and Related Factors of 42 Cases with Acute Leukemia in Chinldren
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To explore the dead reasons and related factors of acute leakemia(AL)in children.Methods The clinical data of 42 dead cases of children with AL,who were admitted to Guangzhou Children's Hospital between Jan.1999 and Mar.2007 were reviewed.Nineteen cases were boys,23 cases were girls;the aging from 4 months to 12 years old and 5 months.Of 42 children,21 cases were acute lymphoblastic leukemia(ALL),18 cases were acute myeloid leukemia(AML),3 cases were unclassified AL.Results Ten out of 12 early-dead cases died of bleeding.Among 30 cases who received chemotherapy,4 cases died of bleeding,others of myelosuppression combined with infection,14 of whom died from sepsis,3 cases with fungal infection,1 case with virus infection,4 cases with unknown pathogens.Sixteen children relapsed among those who got remission,half of them relapsed within 1 year since diagnosed.Treatment-related mortality(TRM)was 90%(27 out of 30 cases).The main causes of TRM were infection and hemorrhage.Hyperleukocytosis and bleeding were 2 risk factors related to early death in new-diagnosed children.Conclusions More emphasizes should be put on bleeding tendency during early treatment in children with AL and treatment of infection complications during induction chemotherapy to reduce TRM.
6.Clinical efficacy of sticking-needle acupuncture plus tendon-regulating manipulation in the treatment of acute ankle sprain
Yan ZHAO ; Yun-Zhen ZHAO ; Sheng-Dong YU
Journal of Acupuncture and Tuina Science 2021;19(6):469-474
Objective: To explore the treatment effect of sticking-needle acupuncture plus tendon-regulating manipulation on pain, swelling and joint dysfunction due to acute ankle sprain.Methods: A total of 80 patients with acute ankle sprain were recruited and divided into a control group and a treatment group according to the random number table method, with 40 patients in each group. Both groups received the same conventional interventions. The control group was treated with additional tendon-regulating manipulation, and the treatment group was treated with tendon-regulating manipulation plus sticking-needle acupuncture. The pain, swelling and joint dysfunction of the ankle were observed in both groups before and after 3 d, 7 d and 14 d of treatment. Results: Before treatment, there was no statistically significant difference in the visual analog scale (VAS) score of pain between the two groups (P>0.05). After treatment, the VAS scores of both groups showed a decreasing trend over time, and the differences within the groups were statistically significant (P<0.05). After 3 d, 7 d and 14 d of treatment, the VAS scores in the treatment group were lower than those in the control group, with statistically significant differences (P<0.05). Before treatment, there was no statistically significant difference in the degree of swelling between the two groups (P>0.05). After treatment, the degrees of swelling in both groups showed a decreasing trend over time, and the differences within the groups were statistically significant (P<0.05). After 3 d and 7 d of treatment, the degrees of swelling in the treatment group were lower than those in the control group, with statistically significant differences (P<0.05). There was no statistical difference in the degree of swelling between the two groups after 14 d of treatment (P>0.05). Before treatment, there was no statistically significant difference in the American Orthopedic Foot Ankle Society (AOFAS) ankle-hindfoot scale score between the two groups (P>0.05). After treatment, the AOFAS ankle-hindfoot scale scores of both groups showed an increasing trend over time, and all the differences within the groups were statistically significant (P<0.05). After 3 d, 7 d and 14 d of treatment, the AOFAS ankle-hindfoot scale scores in the treatment group were higher than those in the control group, with statistically significant differences (P<0.05). Conclusion: Based on the routine intervention, the addition of sticking-needle acupuncture and tendon-regulating manipulation was better in alleviating pain and swelling and improving joint function in patients with acute ankle sprain than the addition of tendon-regulating manipulation alone.