1.Cloning, expression, purification and activity assay of Leucyl-tRNA synthetase from Trypanosoma Brucei
Ying YAO ; Guangwei GAO ; Dawei LI
Chinese Journal of Biochemical Pharmaceutics 2010;31(1):14-18
Purpose To clone and express Trypanosoma Leucyl-tRNA synthetase (LeuRS) gene and to complete the purification and activity assay of LeuRS. Methods We cloned the LeuRS gene from T. Brucei genomic DNA,and cloned it into pBS-T and then into the expression vector pET21a( + ) .The expression of T. Brucei LeuRS was carried out in E. coli BL21 (DE3) RIPL host by optimizing the expression conditions. The products are purified by His-Bind affinity column and verified by Western blot . Enzymatic activity was detected by isotope labeling. Results A DNA fragment at length of 3.2 kb was amplified. Both restriction analysis and sequencing analysis proved that recombinant plasmid pET21a( + )/LeuRS was correctly constructed. The expressed product contained about 20% of total somatic soluble protein and reached a purity of 85% after purification . It was verified by the Western blot. Enzyme unit per millilitre of purified products was about 72. Conclusion Here we report the successful clone, expression and purification of LeuRS from Trypanosoma Brucei ( T. Brucei) in bacterial host. In addition, we tested its enzymatic activity by isotope labeling.This work would be helpful to the design and in vitro screening of Trypanosoma LeuRS inhibitors.
2.STUDIES ON PRODUCTION OF EXPOLYSACCHARIDES OF GRIFOLA FRONDOSA IN SACCHARKID SOLUTION
Dezhu LEI ; Dawei GAO ; Shujuan YU ;
Microbiology 1992;0(03):-
Effects of glucose concentration and pH\|control upon production of exopolysaccharides of Grifola frondosa was studied.The results showed that glucose at concentration of 5% was favorable to exopolysaccharides production,and the pellets benefited from the controlled start pH as well as the controlled final pH of 3 5~4 0 to produce exopolysaccharides.Attempt on the exopolysaccharides production with recycled pellets in the saccharoid solution was also made in this work.
3.Giant nasal septalhaemangioma in pregnancy: one case report.
Dawei ZHANG ; Haihai GAO ; Renjie CHEN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(9):851-852
A pregnant lady in her third trimester presented with a rapidly growing right-sided nasal mass associated with epistaxis and nasal obstruction for six months. Examination showed a non tender, protruding mass completely occluding her right nostril. Wide surgical excision was done under anaesthesia. Histopathology revealed capillary haemangioma. In a gravid patient with a rapidly growing intranasal lesion, capillary haemangioma should be considered as a differential diagnosis.
Diagnosis, Differential
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Epistaxis
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Female
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Hemangioma, Capillary
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diagnosis
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pathology
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Humans
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Nasal Cavity
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physiopathology
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Nasal Obstruction
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pathology
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Nose Neoplasms
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diagnosis
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pathology
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Pregnancy
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Pregnancy Complications, Neoplastic
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diagnosis
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pathology
4.Analysis of the Clinicl Characters and the Incidence Features of Neonatal Congenital Hypothy Roidism
Yan GAO ; Minghui WU ; Dawei LIU ; Wenbin ZHAO
Journal of Medical Research 2006;0(05):-
Objective To explore the incidence features of neonatal congenital hypothy roidism in Lianyungang area.Methods The heel blood samples dried on filter paper was collected from newborns after 72 hours.Serum thyroid stimulating hormone(TSH)level was detected by Dissociation enhancement lanthanide fluoroimmuno assay(DELFIA).The cases diagnosed were analyszd for the clinicl characters and the incidence features.Result One hundred and fifty-eighet children with CH were diagnosed and the patients lived in 63 towns(streets)(66.3%).The distribution of CH was diffuse in Lianyungang.There was significant difference in incidence of CH between 2007 to June of 2008 and before 2006.Eighty-two patients were examined by thyroid gland color ultrasound,among which there were sixty-eight patients whoes thyroid glands were normal(82.9%)and fourteen patients were unusal(absent,replacement,hypogenesis)(17.1%).A couple of twins(male)and two embryoes from the same parents were screened for CH.Conclusion There are iodine lack(MUI300?g/L)individual among people.The iodine nutrition condition of pregnant women is one of important factor to influence the TSH level of newborns.Neonatal screeing is the only valid means to find patients with CH.
5.Efficacy of transurenthral resection of muscle-invasive bladder cancer in the elderly
Hong GAO ; Yan LIU ; Zexing WANG ; Aiwu MA ; Dawei CUI
Chinese Journal of Geriatrics 2012;31(7):586-588
Objective To investigate the clinical effects of transurenthral resection of muscleinvasive bladder tumor (TURBT) in senile. Methods This study was clinical prospective random research.46 senile cases with muscle-invasive bladder cancer treated by TURBT was divided into 2 groups:control group and observed group.The observed group was treated with TURBT combined with intravesical instillation of pirarubi 30 mg immediately after TURBT,the bladder wall around tumor was resected thoroughly.The control group underwent radical cystectomy and pelvic lymph node dissection. Results The clinical improvement rate was higher in observed group than control group (85.7% s.63.6%,X2 =5.3008,P=0.021).There was not difference in 3 years recurrence rate between observed group(25.0%) and control group(13.6%) (x2 =S.3008,P>0.05).The 1 year survival rate were 95.8%,90.9% and 3 years survival rate were 79.2%,72.7% in observed group and control group,respectively,and there were no differences between the two groups(X2 =0.262,P=0.609). Conclusions TURBT combined with intravesical instillation of pirarubi can enhance clinical effect and reduce recurrence rate in the treatment of the elderly with muscle-invasive bladder cancer.
6.Effect of the intestinal ischemia/reperfusion injuries on paracrine patterns of BM-MSCs
Dawei LI ; Guangzhou GAO ; Xin LI ; Tao SUN
Clinical Medicine of China 2012;(z1):34-37
Objective To explore the effect of the intestinal ischemia/reperfusion injury on paracrine patterns of BM-MSCs in order to provide evidence for BMSCs repair.Methods BMSCs were isolated,cultured,characterized,and stained.The rat intestine ischemia/reperfusion model (IRI) was established.Intestinal epithelial mucosal extracts were obtained either from IRI rat or normal rat.Those two type extracts were added into the culture medium,and BM-MSCs were cultured with the two conditioned medium respectively.The culture medium was collected at 0 h and 24 h after culturing with intestinal epithelial mucosal extracts.The levels of TGF-ot,VEGF,FGF and IGF-1 in the medium were assayed by ELISA.Results The contents of all the growth factors detected in cultured medium with intestinal extracts from IRI rats were significantly higher than in regular rats (P < 0.05).The levels of TGF-α,VEGF in culture medium containing IRI epithelium extracts were significantly higher than that in the medium containing normal intestinal epithelium extracts (P < 0.05)Conclusion BM-MSCs can secret TGF-α,VEGF,FGF,and IGF-1 in the presence of intestinal extracts from IRI rat in the culture.These factors might stimulate the differentiation and proliferation of intestinal epithelial cells through paracrine mechanism.
7.A clinical-radiologic-pathologic analysis of telangiectatic osteosarcoma
Zhenhua GAO ; Huaifu DENG ; Quanfei MENG ; Junqiang YIN ; Dawei LIU
Chinese Journal of Radiology 2010;44(6):645-649
Objective To study the clinical, radiologic and pathologic characteristics and diagnostic methods of telangiectatic osteosarcoma for further improving the diagnostic ability.Methods The data of 10 patients with histologically proved telangiectatic osteosarcomas were retrospectively reviewed, and the clinical, radiologic and pathologic characteristics were further analysed in combination with the literature.All 10 patients were examined with X-ray and MRI, and 2 patients with CT.Results Telangiectatie osteosarcomas originated from inferior femur in 5 patients, femur neck in 1 patients, superior humerus in 2 patients and superior segment of tibia in 2 patients.The lesions showed osteolytic bone destruction on X-ray films (n = 10) and CT images ( n = 2), with mild bone expansion in 4 patients.The majority of the edge of the destroyed bone areas was unclear but without sclerotic rim.There were Codman's triangle and soft tissue mass in each patient but no obvious neoplastic bone forming.On MRI, all the lesions were mostly or completely constituted by the multiple cysts with periostnal reaction, and several scatteredly smaller liquid-liquid levels were found within cystic cavity in 7 patients.In all 10 cases, there were pathologic hemocoele similar to aneurysmal bone cysts (ABC), but malignant tumor cells and some neoplastic bones were found in cystic walls or septations.Only a small number of neoplastic bone tissue were seen by microscopy in 6 patients.Conclusions The radio-pathologic characteristics of telangiectatic osteosarcoma include the similar imaging findings of ABC, the common growth patterns of malignant tumors,and the pathologic hemocoele, malignant tumor cells within cystic wails or septations.The comprehensive analysis of clinical, radiologic and pathological data may help clinicans to make a correct diagnosis for telangiectatic nsteosareoma.
8.Effects of different doses of Xuebijing injection in treatment of ischemia reperfusion injury in rabbit limbs
Liang ZHANG ; Wei DAI ; Zhiming GAO ; Jihong ZHOU ; Dawei LIU
Chinese Journal of Trauma 2013;29(8):796-799
Objective To investigate effect and mechanism of Xuebijing administration at various concentrations on ischemia reperfusion injury (IRI) of rabbit limbs.Methods Thirty New Zealand rabbits were divided into control group (n =10),Xuebijing group Ⅰ (n =10) and Xuebijing group Ⅱ(n =10) according to random number table.Rabbit models of IRI in lower extremities were established.Each group received corresponding therapy after reperfusion:rabbits in Xuebijing group Ⅰ were firstly administered 4 ml/kg Xuebijing solution and 6 ml/kg isotonic saline; rabbits in Xuebijing group Ⅱ were administered 2 ml/kg Xuebijing solution and 8 ml/kg isotonic saline; rabbits in control group were simply administered 10 ml/kg isotonic saline.Venous blood samples were collected before reperfusion and at 1 h,2 h,4 h after reperfusion to measure coagulation parameters (APTT,Fib,INR and PT) and biochemical items (ALB,LDH and CK).Results APTT in Xuebijing group Ⅰ presented obvious improvement at 1 h and 4 h after reperfusion as compared with control group (P < 0.01).PT in Xuebijing groups Ⅰ and lⅡ was significantly longer after reperfusion than that before reperfusion (P <0.05).Fib level in Xuebijing group Ⅰ was much higher at 4 h after reperfusion than that before reperfusion (P < 0.05).ALB level at 1 hour after reperfusion showed no statistical differences from that before reperfusion in Xuebijing groups Ⅰ and Ⅱ (P > 0.05).LDH and CK levels in Xuebijing group Ⅰ were much lower than those in control group after reperfusion (P < 0.05).Conclusions Xuebijing injection relieves limb IRI,with better effect in Xuebijing group Ⅰ than in Xuebijing group Ⅱ.Therapeutic mechanism may be associated with its involvement in adjusting clotting function and mitigating injury of muscle tissues.
9.Compare the analgesic effect of intercostal nerve freezing and controlled intravenous analgesia for thoracotomy patients
Yuehua DONG ; Yulei WEI ; Dawei WANG ; Yanjun YANG ; Yongshan GAO
Chongqing Medicine 2014;(15):1866-1867,1870
Objective To compare the clinical efficacy of postoperative analgesia between intercostal nerve freezing and con‐trolled intravenous analgesia in patients of thoracic surgery .Methods 80 patients of thoracic surgery from January 2012 to June 2013 were randomly divided into two groups :Intercostal nerve cryotherapy group (frozen group n=40) and intravenous analgesia group(control group n=40) .Frozen group :the intercostal incision and down each one intercostal and chest tube placement of inter‐costal nerve roots were frozen before sternal closure ;control group :intravenous analgesia pump were used postoperative .According to VAS method to evaluate pain level and observe adverse reactions ,complications and analgesic drug usage of postoperative pa‐tients .Results The analgesic effect of frozen group was better than that of control group within five days after thoracotomy .Com‐pared with the control group ,the incidence of adverse reactions ,postoperative complications ,and analgesic drug usage was signifi‐cantly reduced in frozen group ,there was a significant difference between the two groups (P<0 .05) .Postoperative follow‐up dis‐play :intercostal nerve area in some patients may appear numbness ,dysesthesia ,etc .,but the above situation can return to normal gradually .Conclusion The analgesic effect of intercostal nerve cryotherapy for thoracotomy patients is excellent ,and with few side effects and good safety ,and it is worthy of promotion .
10.Comparison of the effects between the Xuebijing and the Danshen injection on ischemic-reperfusion injury of rabbit limb
Liang ZHANG ; Wei DAI ; Zhiming GAO ; Jihong ZHOU ; Dawei LIU
Chongqing Medicine 2013;(31):3724-3727,3731
Objective To study and compare the effects and mechanisms between the Xuebijing injection and Danshen injection on ischemic-reperfusion injury of rabbit limb .Methods 30 New Zealand rabbits were randomly divided into control group ,Xuebi-jing group and Danshen group ,with 10 in each group .The Crinnion model of ischemic-reperfusion injury of rabbit limb was used . Each group was treated correspondingly after the blood flow was restored .Xuebijing group and Danshen group were treated with 4 mL/kg Xuebijing injection 2 mL/kg Danshen injection in saline with 10 mL total volume .Control group was treated with 10 mL sa-line .Before releasing clip and 1 h ,2 h and 4 h after reperfusion ,the blood samples were collected for detecting clotting function (APTT ,FIB ,PT-INR ,PT) ,biochemistry items (ALB ,LDH ,CK) ,MDA level and SOD level .Results Compared with control group ,the APTT of Xuebijing group in 1 h and 2 h and Danshen group in 4 h after reperfusion improved significantly (P<0 .05) . The PT of Xuebijing group after reperfusion extended significantly compared with the PT before reperfusion (P<0 .05) .The FIB level of Xuebijing group and Danshen group in 4 h after reperfusion was much higher than before (P<0 .05) .The LDH and CK level of Xuebijing group after reperfusion was much lower than that of control group (P< 0 .05) .The MDA level of Xuebijing group in 2 h and Danshen group in 1 h ,2 h after reperfusion was much lower than that of control group (P<0 .05) .The SOD activ-ity of Xuebijing group in 1-4 h and Danshen group in 2 h after reperfusion was much higher than that of control group (P<0 .05) .Conclusion Xuebijing injection and Danshen injection have the relieving effect on the limb ischemic-reperfusion injury by ad-justing clotting function and decreasing free oxygen radicals .In terms of relieving the injury of muscle tissue ,the effect of Xuebijing injection might be better than Danshen .