2.Tolerance to Low Temperature of Transgenic Aloe Plants with TaDREB Gene from Wheat
Hua ZHAO ; Jin ZHAO ; Yinmao DONG ; Congfen HE ; Qin ZHONG
China Biotechnology 2006;0(09):-
Plant expression cassette for TaDREB from wheat was constructed into plasmid pBIR1.Aloe stems were used as explants for the transformation mediated by Agrobaterium.Infected tissues were selected using G418 to generate transformants.In total,58 resistant plantlets to the antibiotics were obtained from the infected explants.The designed primers according to the selective gene npt II and the target gene TaDREB were used to analyze all of the G418 resistant plantlets.PCR results demonstrated that TaDREB were successful transferred into aloe genomic with the transformation efficiency of 0.5%.The transgenic aloe plants were treated under 4℃ for two weeks and then at-20℃ for 30min.The treatment showed that the leaves of negative plants appeared severe evidence of freeze injury with brown,withered and translucent,while the positive plants appeared good growing condition.The activities of enzymes such as peroxidase(POD)and superoxide dismutase(SOD)of transgenic plants which were stressed for 14 days under low temperature were analyzed.The results indicated that the trend of SOD and POD activities in transgenic plants was down-up-up-up,and that in non-transgenic plants was down-up-down-down.The average value of relative electrical conductivity in the positive plants was 0.456 which was lower than 0.685 in the negative plants.It is supposed that transformation of the kind of gene could improve the resistant ability of aloe to low temperature.
3.Dysfunction of epidermal barrier in psoriasis
Hua GU ; Na LI ; Ying TU ; Qin PANG ; Li HE
Chinese Journal of Dermatology 2012;45(2):134-135
ObjectiveTo seek experimental evidence of epidermal barrier dysfunction in psoriasis,and to provide a basis for adjuvant therapy of psoriasis.MethodsPhysiometric methods were used to determine the value of sebum content,transepidermal water loss(TEWL) and water content of stratum corneum in 60 patients with psoriasis and 48 normal human controls.The ultrastructure of lamellar bodies was observed with transmission electron microscopy,and the expression of acid ceramidase in normal skin and psoriatic lesions was detected by using immunohistochemical techniques.ResultsCompared with the normal skin,TEWL value was increased(P < 0.01),but water content of stratum corneum decreased(P < 0.01 ) in psoriatic lesions,and sebum content was similar between normal skin and psoriatic lesions.As electron microscopy showed,lamellar bodies in keratinocytes were reduced in number with a disorganized arrangement and irregular size in psoriatic lesions.The expression of acid ceramidase also decreased in psoriatic epidermis.Conclusions The function of epidermal barrier in psoriasis is impaired,and to restore epidermal barrier function and enhance hydration may serve as an important adjuvant therapy of psoriasis.
4.Effects of Prinsepia utilis Royle oil on the synthesis of ceramide and expression of ceramidase
Ying TU ; Hua GU ; Na LI ; Qin PANG ; Li HE
Chinese Journal of Dermatology 2012;45(10):718-722
Objective To evaluate the effects of Prinsepia utilis Royle oil (PURO) on the synthesis of ceramide and expression of acid ceramidase N-acylsphingosine amidohydrolase 1 (ASH1),and to explore the mechanisms underlying its moisturizing and skin barrier-repairing effects.Methods Keratinocytes from human foreskin tissue were classified into 2 groups to be cultured in keratinocyte-serum free medium (K-SFM) with or without the presence of PURO.Enzyme linked immunosorbent assay (ELISA) was performed to measure the level of ceramide in the culture supernatant of keratinocytes at 0,3,8,24 and 48 hours.The back of nude mice was divided into 4 areas,i.e.,test area,matrix area,blank control area and negative control area.Acetone and ether were used to destroy the epidermal barrier in the test,matrix,and blank control areas,then,the former 2 areas were topically treated with emulsions containing 1% PURO and matrix,respectively,and the blank control area remained untreated.The epidermal barrier remained intact and untreated in the negative control area.Noninvasive methods were used to determine transepidermal water loss (TEWL),epidermal moisture content and skin lipid content in these areas on day 0,1,3,and 7.Skin tissue was obtained from these areas on day 0 and 7 followed by an immunohistochemical study for the quantification of ASH1 expression.Results The level of supernatant ceramide increased with time in the PURO-treated keratinocytes,which was significantly higher at 24 hours and 48 hours than at 0 hour (1.3817 ± 0.100 and 1.3737 ± 0.047 vs.0.7630 ± 0.143,both P < 0.05).The supernatant ceramide was also elevated in the PURO-treated keratinocytes compared with untreated keratinocytes at 24 and 48 hours (both P < 0.05).Noninvasive skin tests showed a gradual decrease in the TEWL,but an increase in the epidermal moisture content and skin lipid content with time in the 3 epidermal barrier-destroyed areas.As far as the test area was concerned,TEWL value was significantly lower on day 3 and 7 than on day 0 (10.85 ± 0.64 and 8.01 ± 0.58 vs.12.65 ± 0.71,both P < 0.05),while a significant increment was observed in the skin lipid content on day 3 and 7 compared with day 0 (29.14 ± 0.40 and 31.30 ± 0.88 vs.27.02 ± 0.65,both P < 0.05),as well as in the epidermal moisture content on day 1,3 and 7 compared with day 0 (13.98 ± 0.28,15.00 ± 0.38 and 15.86 ± 0.18 vs.11.74 ± 0.62,all P< 0.05).On day 7,there was a statistical decline in TEWL value,but an elevation in epidermal moisture content,skin lipid content and ASH1 expression in the test area compared with the matrix area and blank control area (all P < 0.05).Also,the expression of ASH1 was upregulated on day 7 compared with day 0 in the 3 barrier-destroyed areas (all P < 0.05).Conclusion PURO may exert skin-moisturizing and barrier-repairing effects by enhancing the synthesis of ceramide and expression of acid ceramidase ASH1.
5.Innovating Experimental Teaching System and Improving Students’ Practicing Ability——Experimental Teaching Reformation of Environmental Microbiology
Bao-Yan HE ; Hua YIN ; Jin-Shao YE ; Hua-Ming QIN ; Li-Li WANG ;
Microbiology 2008;0(10):-
A new experiment curriculum system of environmental microbiology was established centering on applied microbiology in environmental protection field and emphasizing on design and research experi- ments to motivate the students’ interests for the course, which helped them to improve their ability of think- ing independently and creatively as well as their practicing ability.
6.Assessment of the curative effects and the application value of the radio-frequency ablation in treating uterine leiomyomas by contrast-enhanced ultrasonography
Guangbin HE ; Xiaodong ZHOU ; Qiuyang LI ; Xin MENG ; Miaomiao ZHANG ; Hua HE ; Haiying QIN ; Li WANG
Chinese Journal of Ultrasonography 2008;17(9):793-795
Objective To assess the value of contrast-enhanced ultrasonography(CEUS)in evaluating the therapeutic efficacy of uterine leiomyoma treated by radiofrequency ablation(RFA)and to discuss the value of its application.Methods Forty-four patients which were diagnosed as uterine leiomyomas by ultrasonic guidance needle puncture biopsy preoperation were examined by CEUS before and one week after RFA treatment respectively,and the patho-results were got again after the surgery.Routine ultrasonography was performed in 1,3,6,12 months after the treatment to compare the volume change of uterine leiomyomas with the change of the clinical symptoms.Results After the treatment,CEUS showed that the contrast agent didn't appear in thirty-five patients'uterine leiomyomas and the nonperfusion regions matched the size of the uterine leiomyomas completely.The contrast agent enhanced on the edge of the uterine leiomyomas in nine,the patho-results confirmed that the nonperfusion regions of the contrast agent were necrosis tissues and the marginal local enhancement regions were incomplete necrosis.Three month after RFA treatment,the uterine leiomyomas began to shrink,which showed obviously in 6 months after the treatment.There were significant differences in the parameters before and after the treatment(P<0.05).Clinical symptoms improved obviously in thirty-five patients(90%)after the treatment.Conclusions RFA is an effective minimally invasive surgery of the uterine leiomyoma.CEUS can identify the treatment range and the remaining tissues.It has significant value in guiding the supplementary treatment.
7.Evaluation of the rapid trehalose test for the identification of the Candida glabrata
Chun HE ; Qin LIN ; Zhenying BAO ; Yanhong MENG ; Xiaomei QI ; Hong HUA ; Zhimin YAN
Journal of Peking University(Health Sciences) 2015;(1):186-190
Objective:To explore a rapid and cost-effective method for identification of Candida gla-brata through the comparison of two different methods , using molecular methods of sequencing as gold standard.Methods:From our clinic, 200 strains of suspected Candida glabrata were collected during the last 3 years and selected after incubation in CHROMagar Candida medium for 48 h.By comparing the results of the CHROMagar Candida medium, the identification of the rapid trehalose test for different kinds of strains were analyzed under incubation in the tubes for 3 h, 6 h, and 24 h at 37 ℃and 42 ℃, respectively .All the strains were identified to species level by methods of sequencing .The optimal time and temperature of the trehalose test for the identification of Candida glabrata were assessed .Two different methods, CHROMagar Candida medium and the rapid trehalose test , in identification of Candi-da glabrata were compared.Results:In all the 200 strains, Candida glabrata ferment trehalose with 3 h incubation under 42 ℃ were the optimal time and temperature for fermenting trehalose .The accuracy , sensitivity, and specificity of the rapid trehalose test were 99.00% (198/200), 98.66% (147/149) and 100.00%(51/51).The accuracy rate of CHROMagar Candida medium was 79.50%(159/200), the sensitivity and specificity were only 89.93%(134/149) and 49.02%(25/51), however, compared with the domestic current popular methods , the rapid trehalose test had better time efficiency ratio .Con-clusion:The evaluation results suggest that the rapid trehalose test has advantages in terms of operational convenience and low cost , and the results can be obtained in 3 h.Therefore, it has application value in clinical laboratory .
8.Correction of prominent mandibular angle with aid of rapid prototyping technique
Chuan CAO ; Duo XU ; Shirong LI ; Xia DAI ; Sheng BI ; Hua CHEN ; Xia QIN ; Peng HE
Journal of Third Military Medical University 1983;0(03):-
Objective To investigate the application of computer-aided 3D reconstruction and rapid prototyping(RP) technique in the correction of prominent mandibular angle.Methods Computer tomography scanning and 3D reconstruction were performed on 15 square face patients with prominent mandibular angles,then their actual mandible models were made by RP techniques.Surgical programs were made according to the model,including partial mandibular angle osteotomy,outer mandible table sagittal splitting osteotomy,chin augmentation with autogenous mandibule bones,and so on.In 15 cases,mandibular angle partial cutting was performed in 5 cases,sagittal splitting osteotomy in 6 cases,and mandibular angle partial cutting combined with splitting osteotomy in 4 cases.The autogenous mandibule bones were transplanted for chin augmentation in 3 chin microsomia patients.All the cases were treated according to the position and range set by the RP model.Results All the mandibular models produced by RP techniques were real and complete,which could directly and precisely show the state of the mandible.The operations completed smoothly and accomplished with the expected outcomes designed before operation.In all cases,the width of lower face was efficiently reduced and the face was symmetrical after operation.The follow-up period ranged from 3 months to 1 year in 12 patients,during which their facial appearances were in good condition and the results were satisfactory.Conclusion RP techniques is helpful in precise representation of the state of mandible,which providing ideal surgical models for accurate evaluation of prominent mandibular angle,design of surgical procedures as well as surgery instruction.It can provide good assistance to facial contour plastic surgery.
9.Effects of epidermal proteins and lamellar bodies on epidermal barrier in glucocorticoid-dependent dermatitis
Jue QI ; Hua GU ; Yang TANG ; Ying TU ; Qin PANG ; Li ZHANG ; Li HE
Chinese Journal of Dermatology 2012;45(2):87-90
ObjectiveTo explore the effects of epidermal proteins and lamellar bodies on skin barrier in glucocorticoid-dependent dermatitis.MethodsTotally,60 patients with glucocorticoid-dependent dermatitis and 40 normal human controls were eligible for this study.A noninvasive method using TewameterTM was applied to determine transepidermal water loss (TEWL) value in these subjects.Tissue specimens were obtained from the lesions of 13 patients with glucocorticoid-dependent dermatitis and normal skin of 10 human controls.Subsequently,haematoxylin and eosin(HE) staining was performed to observe the histopathological changes,immunohistochemistry to detect the protein expressions of K6,K10,K14,K15,loricrin,filaggrin,involucrin in epidermis,and electron microscopy(EM) to estimate the density of lamellar bodies in tissue specimens.ResultsCompared with the normal controls,the patients displayed an elevated TEWL value (P < 0.05),which suggested an impaired epidermal barrier.Histopathology of lesions revealed nonspecific inflammatorychanges withmarkeddifferencesbetweendifferentclinicaltypesofglucocorticoid-dependentdermatitis.Immunohistochemistry revealed an attenuated expression of K10,K14,loricrin,filaggrin,involucrin and abnormal expression of K15 in lesional epidermis compared with the normal epidermis (all P < 0.05),hinting a suppression of epidermal differentiation and proliferation as well as an impairment of cornified envelope structure.The number and density of lamellar bodies were also reduced in lesional epidermis compared with the control epidermis.ConclusionsCompared with normal skin,the structure of skin barrier is impaired in lesions of glucocorticoid-dependent dermatitis,to restore skin barrier is essential for the treatment of this entity.
10.Invasive Fungal Infection after Allogeneic Hematopoietic Stem Cell Transplantation in Children
hua-ying, LIU ; chun-fu, LI ; yue-lin, HE ; xiao-qin, FENG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the incidence,clinical status,risk factors and outcomes of invasive fungal infections(IFIs)after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) in pediatric patients.Methods Forty-one Patients who were underwent Allo-HSCT were selected from 2005 to 2006. Of 41 patients, 24 were boys and 17 were girls,aged 2-13 years old. Twenty-six cases with ?-thalassemia, 1 case with adrenoleukodystrophy,and the left 14 cases with other hematologic disorders.Twenty patients underwent bone marrow transplantation,19 patients underwent peripheral blood stem cell transplantation,2 patients underwent bone marrow transplantation and cord blood transplantation.Fourteen patients received Allo-HSCT from HLA-matched sibling donors or HLA mis-matched parents, 27 patients received Allo-HSCT from unrelated donors. Based on different types of transplant, patients were conditioned with busulfan, cyclophosphamide and Anti-thymocyte immune globulin. Fludalabine, total body irration, thiotepa or melphalan was used additionly in some cases. Cyclosporine A and mycophnolate mofetil were used as prophylaxis of graft versus host disease (GVHD).Results IFIs was observed in 5 cases(5/41 cases,12.2%),this comprised cases of proven,probable and possible IFIs at rates of 2.4%,4.9%,4.9%.The time of IFIs was 9-120 d after transplantation,the majority of IFIs in 3/5 cases(60%)children occurred within the first month.The difference of IFIs between patients who recived high-dose corticosteroid and those with no or conventional-dose corticosteroid was significant(?2=8.201 P=0.004);Regarding conditioning regimens,the IFIs of patients who with Thiotepa (TT) was significanthy higher than that of compared with those without TT(?2=9.549 P=0.002).The total effective rate was 40%.The effective rates of the patients with confimed diagnosis,cli-nical diagnosis,and with recommended diagnosis respectively were 100%,0 and 50% respectively.Conclusions IFIs is an important complication after Allo-HSCT,and the high-dose corticosteroid therapy and conditioning regimens with TT are the risky factors for IFIs.Aspergillus is the main pathogen bacteria.