1.Mitochondrial DNA point mutations studies in hereditary ataxia
Chinese Journal of Practical Internal Medicine 2001;0(10):-
Objective To study the possible relationship between mitochondrial DNA(mtDNA)and hereditary ataxia(HA).Methods Polymerase chain reaction(PCR)was used to amplify 4 mtDNA segments of 30 patients with HA、some of their relatives and 35 volunteers.The point 3243、8993、8344 and point 11778 lied in the 4 mtDNA segments respectively.For the PCR products of point 3243 and 8993,restriction fragment length polymophism(RFLP)was performed to search for A3243G、T8993G or T8993C point mutations.For point 8344 and 11778 PCR products,single strand conformation polymorphism(SSCP)was executed to detect mutations.The HA patients’ results of SSCP were compared with their relatives and volunteers’.Sequencing would be carried out to find out exact mutations in those subjects whose SSCP results were abnormal.Results We had not found the A3243G、T8993G or T8993C point mutations in our study.All subjects’ mtDNA segments of point 8344 had not been found mutations.However,a new mtDNA point mutation-A11893G-was identified in 2 patients and 1 relative without symptoms from pedigree 1.Conclusion This new point mutation of mtDNA might be related to HA.
2.Effect of acupuncture on anxiety mood and the expression of β-EP in peraqueductal grey of herion withdrawal rats
Huihua WANG ; Guishu PAN ; Xinyong ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2009;18(6):535-537
Objective To explore the effect of Electroacupuncture on anxiety mood and periaqueductal grey (PAG) β-endorphine expression in rats with morphine abstinence syndrome.Methods The model of heroin-addicted rat was firmly established by the subcutaneous injection of gradually-increased dosages of heroin. Rats were randomly divided into control group, herion-addict group, withdrawal group, electroacupuncture group.Anxiety mood of experimental rats were assessed by Elevated Plus Maze test. Expression of β-EP in PAG was detected by immunohistochemstry. The effect of electroacupuncture at bilateral zusanli and sanyinjiao on anxiety mood and expression of β-EP in PAG was observed in rats with herion abstinence symptom.Results The anxiety index in rats with herion abstinence symptom:the value of OT%,OE%and head-dips times was (12.5±4.3) %, (17.1±6.7)%and (5.7±2.0) times respectively,lower than that in control group [ (26.8±8.7) %,(32.4±6.0) % and (12.2±4.0) times respectively],there existed significant difference(The value of P was 0.003,0.018 and 0.003 respectively).Meanwhile,The expression of β-EP-positive neurons(206.1±23.1)was significantly higher than the control(186.2±15.3). There existed significant difference( P =0.041).Electroacupuncture could increase OT%,OE%,head-dips times[(26.5±8.7)%、(31.8±7.7)% and (9.9±3.1) times respectively] in rats with herion abstinence symptom,which comparied with control group were not statistically distinctive(The value of P was 0.920,0.816 and 0.122 respectively).The expression of β-EP(185.3±11.4)in PAG increased, which comparied with control group(186.2±15.3)were not statistically distinctive( P =0.891).Conclusion Electroacupuncture could inhibit the abstinence syndrome effectively.And there is obvious promotion of β-EP expression in PAG. This is probably one of underlying mechanisms for electroacupuncture improving withdrawal symptoms.
3.Relationship between ionization and the expression of HIF-1α and P53 in anoxia condition
Zhichao FU ; Huihua CHENG ; Fengmei WANG
Journal of Chinese Physician 2009;11(2):188-191
Objective To investigate the effect of ionization to the expression of HIF-1α and p53 of nasopharyngeal carcinoma (NPC) in anoxia condition. Method Nasopharyngeal carcinoma cell line CNE-1 was divided into three groups, control group, radiation group, and hypoxia plus radiation group. The cell viability was analyzed by MTT method, Immunofluorescent technique was performed to de-termine the expression location of HIF-1α in CNE-1. And the expression of HIF-1α and P53 protein were measured with flow cytometry. Re-sults The results of MTr detection showed that the survival fraction of the control group was the highest. The minimum survival fraction was found in the radiation group (P<0.05). There was no HIF-1αprotein expression in cytoplasm except the radiation plus hypoxia group. With the flow cytometer technique, the expression of HIF-1αprotein in the radiation plus hypoxia group was higher than that in the radiation group. No statistical significance was found between the radiation group and the control group. When detecting the expression of P53 protein with the same technique, the expression of P53 was highest in the hypoxia plus radiation group and lowest in the control group (P<0.05). Conclusion The expression of HIF-1α inducted by Hypoxia can prevent NPC cell from the damages of radiation. The HIF-1α decreases the radiosensitivlty through inducing p53 expression in NPC CNE-1 cell line.
4.Study of mitochondrial DNA point mutations at positions 3243, 8993 in hereditary ataxia
Jing WANG ; Huihua LIU ; Shuguang LUO
Journal of Clinical Neurology 1988;0(02):-
Objective To study the possible relationship between mitochondrial DNA point mutations and hereditary ataxia (HA). Methods Polymerase chain reaction (PCR), restriction fragment length polymophism (RFLP) were performed to search A3243G, T8993G or T8993C point mutations in the amplified mitochondrial DNA of extract human perpheral white blood cells of 26 patients with HA and 35 normal controls. Results No point mutations of mitochondrial DNA A3243G, T8993G or T8993C were found in HA group and control group.Conclusion mitochondrial DNA A3243G, T8993G and T8993C mutations are not likely to be genetic factors of hereditary ataxia.
5.Protective effect of ulinastatin on the renal ischemia/reperfusion injury in rats
Congcong CHEN ; Ziming LIU ; Huihua WANG
Chinese Journal of Anesthesiology 1997;0(11):-
Objective To investigate the effects of ulinastatin on renal ischemia/reperfusion (I/R) in rats and assess the possible mechanism. Methods Seventy-five male SD rats weighing 210-250 g were randomly divided into three groups with 25 animals in each group : (Ⅰ) control group received sham operation; (Ⅱ)renal I/R group and (Ⅲ) ulinastatin + I/R group. The animals were anesthetized with intraperitoneal ketamine 50 mg?kg-1 . Bilateral kidneys were exposed through midline incision and bilateral renal pedicles were occluded with atraumatic mini-clamp. The kidneys turned pale. In control group the kidneys were exposed but their pedicles were not clamped, while in ulinastatin group (Ⅲ) ulinastatin 12 500 u was given i. v. 30 min before ischemia and at the beginning of reperfusion. Blood samples and kidneys were obtained at 0, 2, 6, 12, 24 h of reperfusion for determination of blood urea nitrogen (BUN) and serum creatinine (Cr) concentrations and the expression of heat shock protein 70 (HSP 70) and bcl-2 in kidney and for microscopic examination. Paller scoring system was used to evaluate the severity of histological changes ( the higher the score, the worse is the change) .Results BUN and serum Cr levels were significantly lower at 12 and 24 h of reperfusion in group Ⅲ than in group Ⅱ (P 0.05) . Conclusion Ulinastatin can reduce the risk of renal dysfunction and injury associated with renal I/ R. The protective effect of ulinastatin may be associated with the up-regulation of bcl-2 expression.
6.Effect of outpatient nursing intervention in patients with chronic rhinosinusitis
Shan WANG ; Huihua YOU ; Lijun YING
Chinese Journal of Practical Nursing 2017;33(5):327-330
Objective To investigate the effect of outpatient nursing intervention in patients with chronic rhinosinusitis(CRS)on the aspects of clinical efficacy, quality of life and nursing satisfaction after medical therapy. Methods The clinical efficacy according to the judgment standard for clinical effect was valued. Medical outcome study short-form 36-items health survey (SF-36) was used to assess the quali?ty of life of the subjects. Satisfaction Questionnaire was used to evaluate the satisfaction scale of patients. Eighty subjects with CRS were randomly divided into the control group (40 cases) and the trial group (40 cases). The trial group was given nursing intervention on basis of usual nursing. In three months after treat?ment, the data and statistics analysis was purely descriptive by SPSS 20.0. Results Clinical efficacy:in the group, nineteen subjects were cured,fourteen subjects were improved and inefficacy occurred in seven subjects. The effective rate was 77.5%(31/40). In the trial group, twenty-six subjects were cured,twelve subjects were improved and inefficacy occurred in two subjects. The effective rate was 95.0%(38/40). There were differences in general data between the two groups (χ2=5.165,P=0.023). Quality of life:the average score of the control group was 58.10±5.04, while the average score of the trial group was 72.22±5.63, the average score of quality of life in the trial group was significantly higher than that in the control group(t=3.08,P=0.025). Nursing satisfaction:there were significant differences in nursing satisfaction between the control group and the trial group (u=-7.048,P=0.000). Conclusions Outpatient nursing intervention can improve the effect of patients with chronic nasal sinusitis after medical therapy, improve the quality of life and improve nursing satisfaction.
7.Research progress in radiation-induced organ fibrosis
Yanfeng WANG ; Huihua CHEN ; Hang SU
Military Medical Sciences 2016;(1):74-78
Radiation-induced organ fibrosis is a common complication of tumor radiotherapy and radiation accident, which poses a serious threat to patients′life.TGF-β, CTGF, PDGF-BB and NF-κB all play important roles in this pathological process, and drugs targeting these key molecules are also being developed, some of which have shown positive therapeutic effects.This review focuses on the molecular mechanisms and therapeutic progress involved in radiation-induced organ fibrosis, intended to provide a new clue to the complex process of radiation-induced organ fibrosis, and to provide a new strategy for treatment.
8.Factors and mechanism influencing elastin-like polypeptides self-assembled into micron-sized particles.
Huihua GE ; Wenyan WANG ; Guangya ZHANG ; Shibin WANG
Chinese Journal of Biotechnology 2014;30(8):1274-1282
Many factors influence the elastin-like polypeptides (ELPs) self-assembled into micron-sized particles. However, few efforts were made to investigate these factors. Using the ELPs [KV8F]n as the target, we studied systematically the factors with the dynamic light scattering. Our results show that the particle size increased and the uniform of particles decreased with the increase of the molecular weight. The analysis of size variation in self-assembled ELPs in response to changes in salt concentration indicated that the size increased with increasing the salt concentration, and the opposite response was observed when the concentration was above 0.4 mol/L. Under these conditions, the particles are micron-sized and larger than 1.1 μm. However, when the fusions containing the same ELPs and xylanase or 1,3-propanediol oxidoreductase, the size of the self-assembled ELPs particles decreased dramatically, which was only about 1/10 of that of the free ELPs. We proposed that the solvent accessible charged area of the enzymes could interact with the ELPs, the sterical hindrance of the enzymes prevent the aggregation of the ELPs. This might be the most important parameter in altering the particle size sharply.
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9.A new index in evaluating the embolization degree of aneurysms
Jun WANG ; Yan ZHANG ; Huihua ZONG ; Qiaoyu LI
Chinese Journal of Postgraduates of Medicine 2012;35(20):19-21
ObjectiveTo investigate the effect of volume embolization ratio(VER) in evaluating the embolization efficacy of aneurysms.MethodsFifty-three patients who received intracranial aneurysm embolization surgery were all treated with detachable coils endovascular embolization,While fifty-seven intracranial aneurysms were treated.The VER was calculated and the relationships between the VER values and the size,the neck,embolization degree of intracranial aneurysm were analyzed.ResultsThe VER of small,medium,large,and giant aneurysms were (40.8 ± 26.5 )%,( 18.6 ± 16.1 )%,(2.3 ± 2.1 )%,(0.4 ±0.1 )%,respectively,which had statistical significance (F =7.091,P < 0.01 ).The VER of wide-necked and narrow-necked aneurysms were (27.5 ± 23.1 )%,(29.4 ± 26.6)%,which had no statistical significance(P >0.05 ).The VER of complete embolization,neck residual and partial embolization were (41.8 ± 29.3 )%,(31.4 ± 21.2)%,( 12.3 ± 15.1 )%,respectively,which had statistical differences (F =7.970,P < 0.01 ).ConclusionsThe VER is an objective index to evaluate the embolization degree of aneurysms.It has certain prediction significance for the efficacy judgement of embolization of aneurysms.
10.Expression of stromal cell-derived factor-1 in endometriosis by steroid receptor coactivatorson steroid hormone
Xiu SHI ; Wei XU ; Ying SUN ; Huihua DAI ; Xiuli WANG
Chinese Journal of Obstetrics and Gynecology 2014;49(2):114-119
Objectives To study the expression patterns of steroid receptor coactivators (SRC) and steroid-induced stromal cell-derived factor-1 (SDF-1) in endometriosis,and to explore the roles of SRC in the steroid-induced SDF-1 expression endometriosis.Methods From May 2010 to October 2012,16 endometriosis cases at stages Ⅲ or Ⅳ according to the revised American Society for Reproductive Medicine classification undergoing surgery in the First Affiliated Hospital to Nanjing Medical University were enrolled in this study.Their ectopic endometrium were from ovarian endometriomata which were identified pathologically with 9 cases at proliferative phase and 7 cases at secretory phase.The normal endometrium were acquired from the healthy women with normal menstrual cycle (n =10,proliferative phase =5,secretory phase =5).The mnRNA levels of SRC and SDF-1α during the menstrual cycle were detected by quantitative real-time polymerase chain reaction.Ectopic endometrium stromal cells were purified and cultured in medium containing 17β-estradiol (10-8mol/L) or 17β-estradiol (10-8 mol/L) + progesterone (10-6 mol/L).At 24,48,72 and 96 hours,the supernatants were collected to measure SDF-1α expression by ELISA.Ectopic endometrium stromal cells were transfected respectively with siRNA of SRC-1 and SRC-2 using lipofectamine.Two days after transfection,17β-estradiol (10-8 moL/L) or 17β-estradiol (10-8 mol/L) + progesterone (10-6 mol/L) were added into the media.On the third day after the steroid hormones treatment,the media were collected to quantify SDF-1α expression with ELISA.Results (1) Cyclical changes: the SRC-1,SRC-2 and SDF-1 α showed marked cyclic differences in normal endometrium (P < 0.05).In proliferative phase and secretory phase,the SRC-1,SRC-2 and SDF-1 α were 5.6 ± 1.2,3.8 ± 1.1,2.7 ± 0.5 and 2.6 ± 1.0,2.1 ± 1.0,1.6-± 0.5,respectively.There was no periodic variation in the expression of SRC-1,SRC-2 and SDF-1α in ectopic endometrium throughout the menstrual cycle.(2) Steroid-induced SDF-1α expression in ectopic endometrium stromal cells: the 17β-estradiol-induced SDF-1α expression was (1 803 ± 196),(2 272 ± 261) and (2 162 ± 258) ng/L at 48,72 and 96 hours.At the same time points,the SDF-1α expression induced by 17β-estradiol and progesterone was (1 307 ± 150),(1 518 ± 301) and (1 550 ± 144) ng/L,respectively.There was significant difference between two groups (P <0.05).(3) The effects of SRC silencing on steroid hormones-induced SDF-1 α expression in ectopic endometrium stromal cells: the expression of 17β-estradiol-induced SDF-1α at 72 hours was significantly decreased from (2 313 ± 357) ng/L to (1 155 ± 244) ng/L after the silencing of SRC-1 (P < 0.05).After the silencing of SRC-2,the 17β-estradiol-induced SDF-1 α at 72 hours was (1 958 ±324) ng/L.There was no significant difference compared with the before the silencing (P > 0.05).The expression of SDF-1 α at 72 hours induced by 17β-estradiol + progesterone was (1 534 ± 449) ng/L and (2 051 ± 380) ng/L respectively before and after the silencing of SRC-2 and showed the significant difference (P < 0.05).Conclusion During the expression of SDF-1 α regulated by steroids in ectopic endometrium cells,SRC-1 is the major coactivator of 17β-estradiol and SRC-2 is the major coactivator of progesterone.