1.Baculovirus-mediated Expression of p35 Confers Resistance to Apoptosis in Human Embryo Kidney 293 cells
Jian-hua, SONG ; Chang-yong, LIANG ; Xin-wen, CHEN
Virologica Sinica 2007;22(5):389-396
Baculovirus has many advantages as vectors for gene transfer. We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP (Ac-CMV-GFP) could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter. MTT assay suggested that both Ac-CMV-p35 and Ac-CMV-GFP did not have cytotoxic effect on human embryo kidney 293 cells. Cell growth curve showed the Ac-CMV-p35 and Ac- CMV-GFP transduced and non-transduced cells had similar proliferation rate, so baculovirus-mediated p35expression had no adverse effect on cell proliferation. In addition, baculovirus-mediated p35 gene expression protected human embryo kidney 293 cells against apoptosis induced by various apoptosis inducers such as Actinomycin D, UV or serum-free media. These results suggested that the baculovirus vector mediated p35 gene expression was functional and it could be widely used in molecular research and even gene therapy.
2.Preparation and identification of anti-HIV-1 gp120 monoclonal antibody
Yongchen ZHANG ; Jian WEN ; Liang ZHANG ; Lei ZHAO
Chinese Journal of Biochemical Pharmaceutics 2010;31(1):26-28,31
Purpose To Prepare anti- HIV-1 gp120 monoclonal antibodies and to identify the specificity of antibodies in order to provide technique for preparing HIV remedial antibodies. Methods The gene fragment of HIV-1 gp120 was connected to PEGX-4T-2 prokaryotic expressing vector. The vector was cut by enzyme. GST-HIV protein was expressed by E. coli XL1-blue. The BALB/c mice were immunized with purified GST-HIV antigen, and then the fusion of mice spleen cell and myeloma cell SP2/0 was executed as the routine cell-fusion technique. Positive cells were screened by Indirect ELISA. Immuno-blotting assay and Western blot identified the specificity of antibodies. Results External gene section from the recombinant plasmid by sequencing showed the same size of HIV-1 gp120 gene sequences. An external expressed protein band of 32 KD was obtained after purified protein SDS-PAGE electrophoresis. It indicated that six strains of hybridoma cells secreting special monoclone antibodies had been obtained. ELISA results showed that six strains monoclonal antibodies only reacted to HIV-1 gp120 antigen. Western blot results showed that a band with molecular weight 32 kDa was obtained, which could interact with HIV-1 gp120 monoclonal antibody. Conclusion Six strains of hybridoma cells secreting special monoclone antibodies had been obtained. The prepared monoclonal antibodies have established a basis for HIV remedial antibody.
5.Pregnancy outcome in women with rheumatic heart disease
Jian-Hua LIN ; Wan-Wen LING ; A-Juan LIANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(05):-
4.0 cm~2),slight group(2.5-4.0 cm~2),moderate group(1.5-2.5 cm~2)and severe group(
6.Research Progress About Some Problems on Cordyceps militaris
Yan-Feng HAN ; Jian-Dong LIANG ; Wen DU ; Zong-Qi LIANG ;
Microbiology 2008;0(09):-
Cordyceps militaris is of significant economic value in medical care and food exploitation because of its many physiological activities. This paper reviews(1) the taxonomic position of its anamorph,(2) interesting culture ways,(3) strain degeneration and genetic variability,and(4) research progress in bioactive compounds and pharmacological functions.
7.A comparative study on efficiency of different therapeutics methods used for obstructive sleep apnea hypopnea syndrome in children.
Chenyi YU ; Xiaohong CAI ; Zhengwang WEN ; Dongshi LIANG ; Qingqing HU ; Liyan NI ; Jian LIN
Chinese Journal of Pediatrics 2015;53(3):172-177
OBJECTIVETo evaluate effectiveness therapeutic regimens for obstructive sleep apnea hypopnea syndrome (OSAHS) children at an acceptable cost.
METHODThis study was performed at Yuying Children's Hospital of Wenzhou Medical University from Mar. 2008 to Dec. 2010. Prospective random number table method was used for the analysis; 60 children with mild OSAHS were divided into Mild OSAHS Montelukast Treatment (MM) group and Mild OSAHS Adenotonsillectomy Treatment (MAT) group. 32 children in MM group were treated with leukotriene receptor antagonists (LTRAs), while 28 children in MAT group were treated with adenotonsillectomy. Also, 58 children with moderate and severe OSAHS were divided into severe OSAHS Montelukast Treatment (SM) group and severe OSAHS Adenotonsillectomy Treatmen (SAT) group. Twenty-two children in SM group were treated with LTRAs, while 36 children in SAT group were treated with adenotonsillectomy. All selected children were evaluated by polysomnography (PSG) and Obstructive Sleep Apnea-18 (OSA-18) items before and after a six-month treatment. Both records were taken and analyzed, surgical complications and the reason for non-remission after operation were also analyzed. Two therapies were compared based on economic consideration and therapeutic effect. Result (1) PSG: A significant change of a significant change of Apnea Hypopnea Index (AHI) was observed in MM group after the treatment (before receiving the treatment 4.56 ± 1. 26, and after receiving the treatment 3. 48 ± 1. 52, t =3. 50, P <0. 05). But for oxygen desaturation Index (ODI) (MM group 2. 18 ± 2. 19, and MAT group 1. 80 ± 2. 34) and Lowest Oxygen satuation (LSaO2) (MM group 91. 66 ± 2. 34, and MAT group 92. 79 ± 2. 18), there was no significant difference in MM group and MAT group after the treatment (ODI, t =0. 65, and LSaO2 t = - 1. 93, P >0. 05). (2) OSA-18 scores: Significant differences were found in sleeping disorder (before 14. 81 ± 6. 28, and after 10. 56 ± 3. 57), the degree of familial stress (before 13. 56 ± 3. 54, and after 8. 97 ± 2. 96), and OSA-18 total scores (before 52. 66 ± 1. 11, and after 42. 56 6. 48) in MM group after the treatment (sleeping disorder Z - 3. 14, the degree of familial stress Z = -4. 50, and OSA-18 total scores Z= -4. 01, P <0. 05). (3) In addition to the cost of drugs, groups with surgical treatment had a larger economic burden than those with LTRAs treatment. (4) Treatment was totally effective for 28 children (88%) in MM group, and 28 children (100%) in MAT group. Meanwhile, treatment also achieved an obvious effect on 2 children (9%) in SM group, and in 35 children (97%) in SAT group. In MAT group, 3 children improved (11%). And in SAT group, 7 children improved (19%), but treatment was found to be ineffective in 1 case (3%). Among those effective and ineffective cases in groups with surgical treatment, there were 9 children with nasal diseases.
CONCLUSION(1) Surgical treatment is recommended as the first choice for children with moderate and severe OSAHS. And for those who also suffer from nasal diseases, treatment combining drugs with surgery is necessary. (2) LTRAs therapy has a good effect for mild OSAHS. Surgery is also recommended when drugs could not achieve any obvious improvement in clinical symptoms of children with mild OSAHS.
Adenoidectomy ; Blood Gas Analysis ; Child ; Cost of Illness ; Humans ; Oxygen ; Polysomnography ; Prospective Studies ; Sleep Apnea, Obstructive ; surgery ; Sleep Wake Disorders ; Tonsillectomy ; Treatment Outcome
8.Correlation research on contents of podophyllotoxin and total lignans in Sinopodophyllum hexandrum and ecological factors.
Min LI ; Guo-yue ZHONG ; Ao-lin WU ; Shou-wen ZHANG ; Wei JIANG ; Jian LIANG
China Journal of Chinese Materia Medica 2015;40(9):1831-1836
To explore the correlation between the ecological factors and the contents of podophyllotoxin and total lignans in root and rhizome of Sinopodophyllum hexandrum, podophyllotoxin in 87 samples (from 5 provinces) was determined by HPLC and total lignans by UV. A correlation and regression analysis was made by software SPSS 16.0 in combination with ecological factors (terrain, soil and climate). The content determination results showed a great difference between podophyllotoxin and total lignans, attaining 1.001%-6.230% and 5.350%-16.34%, respective. The correlation and regression analysis by SPSS showed a positive linear correlation between their contents, strong positive correlation between their contents, latitude and annual average rainfall within the sampling area, weak negative correlation with pH value and organic material in soil, weaker and stronger positive correlations with soil potassium, weak negative correlation with slope and annual average temperature and weaker positive correlation between the podophyllotoxin content and soil potassium.
Berberidaceae
;
chemistry
;
China
;
Chromatography, High Pressure Liquid
;
Climate
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
Ecosystem
;
Lignans
;
chemistry
;
isolation & purification
;
Podophyllotoxin
;
chemistry
;
isolation & purification
;
Soil
;
chemistry
;
Temperature
9.Transcatheter arterial chemoembolization and delayed surgery for infant hepatoblastoma
xian-liang, HU ; wen-lv, SHEN ; jian-hong, LI ; xue-wu, JIANG ; zhong-xian, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To evaluate the effect of transcatheter arterial chemoembolization (TACE) and delayed surgery for infant hepatoblastoma.Methods TACE was performed with the initial digital subtractive angiography (DSA) under general anesthesia 1-3 times in 8 infants with huge hepatoblastoma, whose age was 2 to 12 months. DSA was done via arterials in hepatoblastoma each time before chemoembolization. The arterials were perfused with chemodrugs and suspensions in ultrasome iodized oil , and were blocked with spring rings. DSA findings indicated that the tumor shrank without new tumorous arterials after 1 month in 6 cases, and 4 of them showed no tumorous staining, and the delayed surgery was performed successfully 1 week later in 6 infants. One boy underwent systemic chemotherapy alone during 6 months after 3 times of TACE. Results TACE therapy did not encounter any major technical problem or toxic reaction caused by chemotherapy. The following DSA test 4 weeks later did not detect any new tumorous vessels in 6 cases. Six children received TACE and surgery had been followed-up with no tumor recurrence for months averagely. The boy underwent TACE and venous chemotherapy for 6 months , without surgery , had been followed-up for 48 months until the present report. CT, AFP and DSA did not show any hints of tumor recurrence. Six cases receiving 3 times TACE combined with surgery survived without tumor recurrence. Conclusions TACE is a very effective, safe and helpful therapy for hepatoblastoma, which stressed the repeated use of spring ring to block tumor vessels lastingly if necessary. If surgery is required, DSA test is needed beforehand to detect new tumorous vessels or neoplasm. If there is any , TACE is repeated. TACE combined with surgery may provide an additional promising choice in the treatment of hepatoblastoma, and repeated TACE alone may cure hepatoblastoma in infants.
10.Evaluation of Blood Gas and Dynamic Monitor of Respiratory Mechanics in Respiratory Failure Children with Mechanical Ventilation
jian-li, CHEN ; li, HUANG ; yan-xia, XU ; li, PENG ; xue-wen, LIANG
Journal of Applied Clinical Pediatrics 2006;0(18):-
Objective To explore the clinical significances of blood gas and dynamic monitor of respiratory mechanics in respiratory failure children with mechanical ventilation.Methods Sixty-four children who need mechanical ventilation over 24 hours were divided into pulmonary disease group(44 cases) and non-pulmonary disease group(20 cases) in pediatric intensive care unit(PICU),from Jan.2005 to May 2008.By Evita 4 ventilator,dynamic monitor of respiratory mechanics,such as respiratory system compliance(Crs),airway resistance(Raw),mean airway pressue(MAP),minute ventilation(MV),oxgen index(OI),Qs/Qt,respiratory index(RI) and blood gas in these children were determined.Their characteristics and clinical significance were analyzed.Results Many parameters such as Crs,Raw,MAP,MV,Qs/Qt,OI,RI were significantly different between 2 groups(Pa