1.Analysis of peblA gene sequence and construction of eukaryotic expression recombinant plasmid of peblA gene of Campylobacter jejuni Pen:19
Hui ZHENG ; Fangcheng CAI ; Bing DENG ; Min ZHONG
Journal of Third Military Medical University 2003;0(07):-
Objective As the genome sequence of Campylobacter jejuni is a hypervariable sequence, the peblA gene sequence of Campylobacter jejuni Pen∶19 was sequenced and the peblA gene conservative was identified. To construct the eukaryotic expression recombinant plasmid of the peblA gene of Campylobacter jejuni Pen∶19 . Methods Total DNA of Campylobacter jejuni Pen∶2, Pen∶8, Pen∶19 and Pen∶21 as templates respectively, peblA gene DNA with KpnⅠ and EcoRⅠ sites was amplified by PCR using the same primer and the PCR product of Pen∶19 was sequenced. Because most close association with Guillain-Barre syndrome, the PCR product of Pen∶19 was selected as target gene and cloned into pcDNA3.1(+) and constructed the recombinant plasmid that was identified by endonuclease digestion and PCR and confirmed by sequencing. The recombinant plasmid was then transfected into mammalian cell COS-7 for expression. The transient expression was investigated by RT-PCR. The expression of peblA gene in the culture supernatant of positive clones was analyzed by ELISA. Results The peblA gene sequence of Campylobacter jejuni Pen∶2 and Pen∶19 was identical. The target gene was amplified from COS-7 cells transfected with recombinant plasmids by RT-PCR. PEBl protein could express in the culture supernatant in the transfected COS-7 cells by ELISA. Conclusion The peblA gene of Campylobacter jejuni was conservative. The recombinant plasmid was constructed and expressed in COS-7 cells successfully. The results obtained lay the foundation for research on development of Campylobacter jejuni DNA vaccine.
2.The study of miR-216a induces cell apoptosis by targeting PKCαin gastric cancer
Xuemei ZHANG ; Hui LAN ; Fang LIU ; Min DENG ; Xiaoqiu LIANG
Chongqing Medicine 2013;(33):4051-4052,4055
Objective To confirm whether miR-216a suppresses cell proliferation and induces cell apoptosis by targeting PKCα, thus to reveal molecular mechanism that miR-216a functions as a tumor suppressor in gastric cancer .Methods PKCα3′untranslat-ed region(UTR)-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-216a on luciferase activity .MGC-803 cells were transfected with miR-216a mimics ,and next Western blotting was performed to detect the expression of PKCαprotein .The effects of PKCαdownregulation on cell proliferation and apoptosis were observed after PKCαsiRNA were transfected into MGC-803 cells .MGC-803 cell proliferation assays were performed when cotransfected with miR-216a mimics .Results The result demonstrated miR-216a could bind to the 3′UTR of PKCαand inhibited the luciferase activi-ty ,cut the 41% .PKCαprotein expressions were significantly down-regulated when miR-216a was overexpressed in MGC-803 .siR-NA-mediated downregulation of PKCα could suppress the potentials of cell proliferation and induce apoptosis .Conclusion miR-216a suppresses cell proliferation and induces apoptosis by targeting PKCαmRNA 3′UTR in gastric cancer .
3.Changes of Activity of Peripheral Blood Mononuclear Cells Nuclear Factor-?B in Children with Primary Nephrotic Syndrome and the Invention of Astragalus
fu, ZHONG ; yan, GAO ; xiao-yuan, ZHAO ; ying-min, DENG ; huan-hui, CHEN
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To investigate the pathogenesis of peripheral blood mononuclear cells(PBMC) nuclear factor kappa-?B(NF-?B) in children with primary nephritic syndrome(PNS) and the effect of astragalus on the activity of NF-?B.Methods Twenty-five children with PNS and 20 normal children were studied.Isolated PBMC were separated from 5 mL venous blood in asepsis condition.NF-?B stimulator,NF-?B inhabitor and astragalus were added into the different tubes of PBMC,respectively.The nuclear protein was extracted from the pellets and the optical density(A) values of nuclear protein was measured by enzyme-linked immunosorbent assay(ELISA).Results The activity of PBMC NF-?B in PNS group was higher than that in normal group(P0.05).Astragalus could decrease the activity of PBMC NF-?B which had been stimulated by interleukin-1?(IL-1?)(P
4.Ultraviolet Mutation Breeding Aspect of the Biological Control of Postharvest Diseases by Kloeckera apiculata in Citrus Fruit
Jia-Yun XIANG ; Bo-Xun DENG ; Yu-Jia LIU ; Hui-Min LIU ; Yan-Xi ZU ; Hui YU ;
Microbiology 1992;0(01):-
A series of experiments were conducted to study the mutation of Kloechera apiculata by many kinds of treatments such as UV and UV+LiCl.The optimal dosage disposal was determined:15 W 30 cm under Ultraviolet irradiation for 20 s,UV+LiCl under Ultraviolet irradiation for 20 s and added LiCl 0.3% (w/v).One strain(UV20-13)which had obvious physiological characteristic was obtained.the incidence of blue and green mold of citrus was reduced by 25.56%and 10.00%in vivo experiment after 7 days respectively. The strain UV20-13 was tested by the experiments of subculture and dynamic growth,and the results showed that the strain UV20-13 was better than K.apiculata in the growth characteristics,and it did not appear retrogression,reversion mutation ect after subculturing 10 generations.Therefore the strain UV20-13 had genetic stability.
5.Therapeutic observation of tuina manipulation for lumbar intervertebral disc herniation
Zhi-Qi ZHANG ; Xia LIU ; Huan ZHONG ; Duo-Xi DENG ; Guo-Min LI ; Jia-Qin DENG ; Zhao-Ping LIU ; Hui XIE ; Xiao-Rong CHANG
Journal of Acupuncture and Tuina Science 2019;17(2):116-123
Objective:To observe the clinical efficacy of balance-impact tuina therapy for lumbar intervertebral disc herniation (LIDH).Methods:A total of 118 eligible LIDH patients were randomized into an observation group and a control group by the random number table method,with 59 cases in each group.The observation group was intervened by balance-impact tuina therapy,while the control group was intervened by conventional tuina therapy,both for consecutive two weeks.The scores of visual analog scale (VAS),Oswestry disability index (ODI),quality of life questionnaire-core 30 (QOL-C30) were observed before and after treatment;the relapse rate was estimated at the sixth month and twelfth month following the treatment.The data were statistically analyzed.Results:After intervention,the total effective rate was 96.6% in the observation group versus 91.5% in the control group,and the between-group difference was statistically significant (P<0.05).The VAS and ODI scores declined significantly after treatment in both groups (all P<0.05),and the observation group was markedly lower than the control group (P<0.05,P<0.01).The QOL-C30 score increased significantly after treatment in both groups (both P<0.05),and the observation group was markedly higher than the control group (P<0.05).The relapse rates at the post-treatment sixth month and twelfth month in the observation group were lower than those in the control group (P<0.05,P<0.01).Conclusion:Compared with the conventional tuina therapy,the balance-impact tuina therapy shows advantage in lessening pain,improving the function and enhancing the quality of life in the treatment of LIDH,and it has a lower relapse rate.Thus,this therapy is worth promoting in clinic.
6.Anatomic and radiological study on posterior pedicle screw fixation in the atlantoaxial vertebrae of children.
Xiong-wei DENG ; Zhi-hai MIN ; Bin LIN ; Fa-hui ZHANG
Chinese Journal of Traumatology 2010;13(4):229-233
OBJECTIVETo investigate the feasibility of posterior fixation with 3.5-mm pedicle screws in the atlantoaxial vertebrae of children.
METHODSIn this study, atlantoaxial vertebrae specimens were obtained from 10 cadavers of children aged 6-8 years. We measured the height and width of the C(1) pedicle and the midportion of C(1) lateral mass; the width of C(1) posterior arch under the vertebral artery groove and the height of the external and internal one-third of this part; the external, internal height and the superior, middle, inferior width of the C(2) pedicle (transverse foramen). Furthermore, computed tomography (CT) axial scan was performed on 20 age-matched volunteers to obtain relative data of their atlantoaxial vertebrae. We measured the length and width of the C(1) and C(2) pedicles in the atlantoaxial cross-sectional plane. On CT workstation, we also measured the angles between the longitudinal axes of the atlantoaxial pedicles and the midsagittal plane.
RESULTSFor the cadaveric specimen group, the height and width of the C1 pedicle were (5.26+/-0.44) mm and (6.26+/-0.75) mm respectively. The height of the medial one-third of the C1 posterior arch under the vertebral artery groove was (4.07+/-0.24) mm. The external, internal height and superior, middle, inferior width of the C2 pedicle was (6.86+/-0.48) mm, (6.67+/-0.49) mm, (6.63+/-0.61) mm, (5.41+/-0.39) mm and (3.71+/-0.30) mm, respectively. For the volunteer group measured by CT scan, the height and width of the C(1) pedicle were (5.47+/-0.34) mm and (6.63+/-0.54) mm respectively, while (6.59+/-0.51) mm and (5.13+/-0.42) mm of the C2 pedicle. The angles between the atlas, axis pedicles and the midsagittal plane were (9.60+/-1.32) degree and (27.80+/-2.22) degree respectively.
CONCLUSIONIt is feasible to place a 3.5-mm pedicle screw in the C(1) and C(2) pedicles of children aged 6-8 years old.
Axis, Cervical Vertebra ; anatomy & histology ; diagnostic imaging ; surgery ; Bone Screws ; Cervical Atlas ; anatomy & histology ; diagnostic imaging ; surgery ; Child ; Humans ; Radiography
7.Clinical utility of reticulocyte hemoglobin content for the diagnosis of iron deficiency anemia in children.
Li-Sha DENG ; Hui-Min TENG ; Yan-Shan LI
Chinese Journal of Contemporary Pediatrics 2011;13(3):212-215
OBJECTIVETo study the clinical utility of measuring reticulocyte hemoglobin content (CHr) in the diagnosis of iron deficiency anemia (IDA) in children.
METHODSOne hundred children with IDA at ages of 1 to 6 years and 50 healthy children were enrolled. Red blood cell parameters, CHr, hemoglobin (Hb), red blood count (RBC) and mean corpusular volume (MCV), were determined using the Blood Cell Analyzer. Serum ferritin (SF) levels were determined using radioimmunoassay double antibody techique. Soluble serum transferrin (sTfR) levels were determined using ELISA.
RESULTSThe values of Hb (100 ± 6 g/L vs 126 ± 8 g/L) and CHr (18 ± 5 pg vs 31 ± 3 pg) in the IDA group were significantly lower than normal controls (P<0.01). SF levels (11 ± 4 μg/L) in the IDA group were also lower than normal controls (59 ± 36 μg/L) (P<0.01). In contrast, the values of sTfR in the IDA group were significantly higher than normal controls (4.8 ± 2.1 mg/L vs 1.4 ± 0.6 mg/L; P<0.01). In both groups, there was a positive correlation between the values of CHr and Hb [r=0.540 (control group), r=0.734 (IDA group); P<0.01]. In the IDA group, CHr was positively correlated with SF(r=0.464; P<0.01) and negatively correlated with sTfR(r=-0.450; P<0.01). When the cut-off value of CHr was 27.8 pg, the sensitivity and specificity for the diagnosis of IDA were 88.0% and 90.0%, respectively and the area under the ROC curve was 0.948.
CONCLUSIONSCHr can be used as an index for the diagnosis of IDA in children.
Anemia, Iron-Deficiency ; blood ; diagnosis ; Child ; Child, Preschool ; Female ; Hemoglobins ; analysis ; Humans ; Infant ; Male ; ROC Curve ; Receptors, Transferrin ; blood ; Reticulocytes ; chemistry
8.Clinical and gene study on one pedigree of hereditary spinocerebellar ataxia type 7
Yan HAN ; Yang-Tai GUAN ; Hui-Min ZHENG ; Su-Ju DING ; Jian-Ming JIANG ; Ben-Qiang DENG ; Tao WU
Chinese Journal of Neurology 2000;0(04):-
Objective To summarize the clinical characteristics and make genetic diagnosis in the patients with hereditary spinocerebellar ataxia type 7 (SCA7).Methods Pedigree analysis and clinical examination were performed in one family with SCA7 by clinical findings,of which retinal morphology and visual electrophysiology were available on part numbers.The polymorphic cytosine adenine guanine (CAG) repeats in the encode region of SCA7 gene were detected by combining polymerase chain reaction with deoxyribonucleic acide (DNA) sequencing on 19 familial numbers and 12 controls.Results 6 patients were identified,who manifesting cerebellar ataxia,decreased visual acuity and colour vision defect,as was pigmentary retinopathy on fundoscopy;The 6 patients had not only extinction of the electroretinogram (ERG) but also remarkably reduced amplitudes of oscillatory potentials and flash-visual evoked potentials. On normal alleles CAG repeat size ranges from 8 to 25 repeats,wherease on mutated alleles of the 6 numbers it ranges from 50 to 97 repeats.The 6 numbers were diagnosised as SCA7 patients.One asymptomatic individual of this family,who displayed a normal allele with 18 CAG repeats and another containing abnormal expantion of 56 repeats,was diagnosised as a asymptomatic carrier whose age maybe still below the age of onset.Conclusion The clinical manifestations of SCA7 are heterogeneous,and the detection of CAG repeats can provide an effective way for the gene diagnosis and the prediction of asymptomatic patients.
10.Experience of Professor JIN Bao-fang in Using Yangjing Decoction in Treatment of Abnormal Sperm Sterility
Guang-Hui CHEN ; Da-Lin SUN ; Bing CHEN ; Xin-Dong ZHANG ; Wei-Min DENG
Chinese Journal of Information on Traditional Chinese Medicine 2018;25(9):107-108
Male sterility affects human reproduction seriously. Modern medicine has not yet fully understood the reasons for abnormal sperm, so clinical treatment is mainly based on experience, but the effect is inaccurate. According to "kidney controls reproduction" and "chronic illness causes blood stasis" theory, Professor JIN Bao-fang treats abnormol speerm sterility by tonifying kidney and activating blood circulation to remove obstruction with modified Yangjing Decoction, which has achieved good efficacy.