1.Cloning for Full-length Pyruvate Orthophosphate Dikinase Gene of Sugarcane
Fu-Ming YANG ; De-Chang XU ; Ai-Ju HOU ;
China Biotechnology 2006;0(10):-
Compared with C3 plant,C4 plant had evident growth advantage,higher rates of water and nutrition using,and higher bio-yield than C3 plant,Sugarcane was one of the typical C4 plants.DNA was extracted from sugarcane leaves,primers were designed by the cDNA sequence of PPDK gene from GenBank.Then DNA was amplified by LA-PCR(Long Acute PCR) method,ligated into pMD18-T vectors,and transformed into E.coli.JM109.Full-length PPDK gene sequence of sugarcane was obtained,the sequence was 13.5kb in length.For convenient of the next experiment,two digest sites(XhoI and NotI) were introduced into primers,the full-length PPDK gene was splitted to two parts,and was ligated into pMD18-T simple vectors,respectively.Whole PPDK gene clone of sugarcane was finished.The strains were storaged in the lab.
2.Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor.
Hui-hua LI ; De-heng YAO ; Jian XU ; Wei WANG ; Qiang CHANG ; Ming-hua SU
China Journal of Chinese Materia Medica 2015;40(9):1693-1698
Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
Biomass
;
Bioreactors
;
Cell Culture Techniques
;
instrumentation
;
methods
;
Culture Media
;
chemistry
;
metabolism
;
Eriobotrya
;
chemistry
;
growth & development
;
metabolism
;
Triterpenes
;
analysis
;
metabolism
3.The tolerance to 188Re-HEDP treatment in patients with bone pain from osseous metastases
Ai-ping, CHENG ; Shao-liang, CHEN ; Wen-guan, LIU ; Xue-fen, CHEN ; Chang-de, XU
Chinese Journal of Nuclear Medicine 2011;31(2):77-81
Objective To study the tolerance to 188Re-1-hydroxy-1 ,1-ethylidene disodium phosphonate(HEDP) in patients with bone pain caused by osseous metastases. Methods Thirty-one patients(10with prostate cancer, 9 with breast cancer, 3 with lung cancer, 5 with liver cancer, 2 with rectal cancer, 1with esophageal cancer and 1 with renal cancer) received a single injection dose of 188Re-HEDP. The patients were divided into four groups according to the injection dose: 20 MBq/kg (6 patients), 30 MBq/kg(6 patients), 40 MBq/kg (9 patients), and 50 MBq/kg (10 patients). Haematological toxicity (WHO grading) of grade Ⅲ- Ⅳ was considered unacceptable. Vital signs and adverse effects after injection were recorded for 8 weeks. Blood counts were measured weekly during a period of 8 weeks. Biochemical parameters and electrocardiogram were assayed at week 4 and 8. Statistical analysis was performed for per-protocol (pp) population (t-test). Results Twenty-seven patients belonged to PP population with 5 in the group of 20 MBq/kg, 5 in the group of 30 MBq/kg, 8 in the group of 40 MBq/kg and 9 in the group of 50 MBq/kg.No obvious adverse effects and no significant change of vital signs, electrocardiogram, liver and renal function were found after injection. Alkaline phosphatase was slightly higher than baseline at week 4 and 8 after therapy, but the difference was not statistically significant. In the 20 MBq/kg group, reversible grade Ⅰ leucopenia was noted in 1 patient. In the 30 MBq/kg group, 2 patients showed reversible grade Ⅰ leucopenia including 1 alone with reversible grade Ⅲ thrombopenia. In the 40 MBq/kg group, reversible grade Ⅰ leucopenia and thrombopenia was observed in 1 patient and reversible grade Ⅱ leucopenia and thrombopenia in another patient. In the .50 MBq/kg group, 3 patients showed reversible grade Ⅱ leucopenia. The lowest level of thrombopenia was at week 4(143.5 × 109/L), leucopenia at week 6 (5.4 × 109/L) and anaemia at week 8(t = 3.1325, 3.3156, 3.4917, all P < 0. 05 compared with baseline). At week 8, the mean level of platelet and leucocyte recovered to baseline. "Bounce pain" was found in 2 of 27 patients (7.41%).Conclusions The dose of 20 MBq/kg, 30 MBq/kg, 40 MBq/kg or 50 MBq/kg of 188Re-HEDP do not cause significant side effects on cancer patients with bone metastases, though there is a tendency that the haematological toxicity may increase as the dose of 188Re-HEDP increases.
4.Analysis of clinical speciality of invasive fungai infection on 137 cases
De-Chang CHEN ; Liang ZHAO ; Xing-Yi YANG ; Zhao-Fen LIN ; Yong-Hua XU ; Chang-Xin GUO ;
Chinese Journal of Emergency Medicine 2006;0(10):-
Objective To analyze the clinical speciality of invasive fungal infection(IFI)and provide doctors with clinical evidence for early anti-fungal therapy.Method One hundred and thirty-seven patients with 91 male and 46 female,who suffered from invasive fungal infection in ICU from January.1,2000 to June 30, 2006,were enrolled in this study.The age ranged from 17 to 82 years old.Out of 137 patients with IFI,the percentage of albicans candida,glabirate candida,tropicalis candida and parapsilosis candida were 47.4%, 26.3%,20.4% and 3.6%,reseparately.The sputum,urine,blood and other drainages were collected to perform the fungal examination after three days of admission every three days.Results Of 137 patients,42 of them were complicated with hemorrhage,53 patients with IFI developed candida anthema in the chest,abdomen and extremity.,49 patients suffering from IFI had organ dysfunction.The chest image revealed that infiltration caused by IFI especially occurred in apex of lung in some patients.The pathogen analysis displayed that albicans candidiasis easily developed candida anthema,glabirate candidiasis frequently resulted in organ dysfunction,and tropicalis candida led to hemorrhage in some organs.Conclusions The clinical specialty,of IFI caused by candida included hemorrhage,candida anthema,organ dysfunction,and infiltration in apex of lung.
5.Clinical study on treatment of advanced non-small-cell lung cancer with Chinese herbal medicine in different stages combined with chemotherapy.
Zhen-ye XU ; Chang-juan JIN ; De-yi SHEN
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(10):874-878
OBJECTIVETo observe the effect of staged Chinese herbal medicinal therapy combined with chemotherapy in treating patients with non-small-cell lung cancer (NSCLC) of stage III or IV.
METHODSAdopting prospective randomized controlled multi-centered method, the 116 patients enrolled were assigned to the treated group (n = 60) and the control group (n = 56). The control group was treated by chemotherapy alone, while the treated group treated by chemotherapy combined with Chinese herbal medicine, i.e. Kangliu Zengxiao Decoction (KLZX) was administered in the chemotherapy stage, followed with Feiyanning Decoction (FYN) in the stage after ending the chemotherapy. The survival time, Karnofsky score, main clinical symptoms and adverse reactions, etc. were observed.
RESULTSThe median survival time was 15.57 months in the treated group, which was higher than that in the control group (11.17 months, P< 0.01). The improvements in tumor related symptoms such as fatigue, dyspnea, etc. in the treated group were better than those in the control group (P <0.05). In addition, adverse reactions such as leucopenia, digestive reaction (such as nausea and vomiting) in the treated group were less than those in the control group (P <0.05).
CONCLUSIONChinese herbal medicine combined with chemotherapy shows favorable effect in improving quality of life and prolonging survival time on patients with advanced NSCLC.
Aged ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Phytotherapy ; Prospective Studies ; Single-Blind Method ; Treatment Outcome
6.Evaluation of bubble oxygen inhalators' performances and an investigation on their solutions for improvement.
Mian-kang CHEN ; Zheng-hai SHEN ; Xun-liang XU ; Jun-cheng BAO ; Chang-shan ZUO ; De-jun TANG ; Jun YANG
Chinese Journal of Medical Instrumentation 2007;31(4):295-296
This paper analyses the defects of bubble oxygen inhalators currently used, and investigates into their solutions for improvement.
Oxygen Inhalation Therapy
;
instrumentation
;
methods
;
Oxygenators
;
standards
7.Enriching of total flavonoids from Herba Leonuri with polyamide and macroporous resin.
Dan-Ni YU ; De-Sheng XU ; Yi FENG ; Hong-Chang WEI
China Journal of Chinese Materia Medica 2008;33(3):264-268
OBJECTIVETo study the enriching method of total flavonoid from Herba Leonuri with polyamide and macroporous resin.
METHODSeven enriching and purifying methods were compared with the yield and purity as indexes. The method of enriching with polyamide and macroporous resin was confirmed and the process of purifying was determined by orthogonal design.
RESULTD101 resin is packed by wet method, the ratio of diameter to height is 1:7. After mixed with the extract liquids, the weight of wet resin increased to 3 times of the dry resin. Evaporated the wet resin to dryness, mixed well with a little of 95% ethanol and dry polyamide powder, evaporated them to dryness again. Elute with deionized water until the effluent being colourless, then loaded it on the macroporous adsorptive resin, elute with 50% ethanol, the volume of effluents was collected to 7 times of the column volume. The purity of total flavonoids reached to 23%, while the diversion rate from raw Herba Leonuri was to 69%.
CONCLUSIONThe process is simple and convenient, and the regeneration of resin is easy, which has a good application foreground.
Absorption ; Flavonoids ; isolation & purification ; Leonurus ; chemistry ; Nylons ; chemistry ; Resins, Plant ; chemistry
8.Overlapping coverage with bilateral shaft based vascularized dartos pedicle in Snodgrass hypospadias repair
Da-Xing TANG ; De-Hua WU ; Chang TAO ; Shui-Heng YAN ; Shan XU ; Yong HUANG ; Cong ZHANG ; Min-ju LI
Chinese Journal of Urology 2001;0(10):-
Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.
9.Isolation and structure determination of steroidal saponin from Dioscorea zingiberensis.
De-ping XU ; Chang-ying HU ; Lin WANG ; Xiao-chun WANG ; Zi-jie PANG
Acta Pharmaceutica Sinica 2009;44(1):56-59
To study the chemical constituents of Dioscorea zingiberensis Wright, the EtOH extract of fresh rhizomes of D. zingiberensis was concentrated and partitioned further to produce petroleum ether-, ethylacetate-, n-butanol- and water-soluble fractions. The water-soluble fraction was subjected to column chromatography on macro resin AB-8, and the final products were obtained by repeated reversed-phase ODS and MCI gel CHP 20P column chromatography. Structures of compounds were elucidated by 1H NMR, 13C NMR, 135DEPT, HMQC, HMBC and TOCSY spectroscopic analyses. A new steroidal saponin was isolated, which was identified as (25R)-26-O-(beta-D-glucopyranosyl)-furost-5-en-3 beta, 16, 20, 26-tetraol-22-seco-3-O-beta-D-glucopyranosyl-(1--> 3)-beta-D-glucopyranosyl-(1--> 4)-[alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside. The compound is a novel skeletally steroidal saponin, named as zingiberenin F (1). It was reported for the first time from D. zingiberensis Wright.
Dioscorea
;
chemistry
;
Molecular Structure
;
Plants, Medicinal
;
chemistry
;
Rhizome
;
chemistry
;
Saponins
;
chemistry
;
isolation & purification
10.Preoperative Doppler evaluation of vascular perforators in the anterolateral thigh flap harvest.
Zhong-fei XU ; Wei-yi DUAN ; De-hao SHANG ; Wei DAI ; Xiao-jiao ZHENG ; Chang-fu SUN
Chinese Journal of Stomatology 2011;46(5):290-292
OBJECTIVETo evaluate the accuracy of Hadeco ES-1000spm hand-held doppler during the anterolateral thigh (ALT) flap harvest.
METHODSTwenty-five patients (26 sides) with ALT flaps for head and neck reconstruction between May 2005 and May 2010 received preoperative Doppler examination for the location of the cutaneous perforators of ALT flaps. The Doppler signals and body mass index (BMI) were recorded preoperatively according to ABC system. The locations of Doppler signals and of the actual cutaneous perforators at surgery were plotted and compared. The diameter of perforators was measured.
RESULTSOne to three cutaneous perforators of the ALT flap were consistently found at specific locations. They were named perforators A, B, C from proximal to distal. Perforators A, B and C were present in 15 (58%), 24 (92%) and 20 (77%) cases and the diameter (> 0.5 mm) of A, B and C were 11/15, 22 (92%) and 8 (40%) respectively. The Doppler signal was within 0.5 cm of the actual perforator location in 85% flaps. The accuracy of Doppler decreased with increase of BMI.
CONCLUSIONSPreoperative assessment by hand-held Doppler is useful in predicting the perforator vessels' locations and diameter although it's accuracy is limited.
Adult ; Aged ; Body Mass Index ; Female ; Head and Neck Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Perforator Flap ; Preoperative Care ; Reconstructive Surgical Procedures ; methods ; Thigh ; blood supply ; diagnostic imaging ; surgery ; Ultrasonography, Doppler