3.CLONING OF NATTOKINASE GENE AND EXPRESSION IN E. COLI
Li-Xin LUO ; Zhi-Li HUANG ; Ru-De YANG ; Jun-Jian LING ; Shi-Zhong LIANG ;
Microbiology 1992;0(03):-
In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.
4.Expression and Identification of LfcinB Gene in Pichia pastoris
Jun-Bo YI ; De-Xin HUANG ; Ling-Yun LI ; Feng LIN ;
Microbiology 1992;0(02):-
In present study,bovine Lactoferricin was first secretly expressed in Pichia pastors yeast expression system.The synthesized LfcinB gene fragment was cloned into expression vector pPIC9K,and then obtained recombinant plasmid,designated as pPIC9K-LfcinB,was linearized and transformed into Pichia pastors strains SMD1168 by electroporation.The transformants were screened with Geneticin and multiply-copy colonies were harvested,in which LfcinB gene was verified to inserted into yeast chromosome stably.The positive recombinant Pichia strains were induced with methanol to express LfcinB in culture supernatant.It's expressive products has high activity of killing bacteria.We concluded that LfcinB gene was cloned and integrated into yeast chromosomes,and obtained expression peptide was tested to have high antibacterial activity.
6.Expression and Antigenic Analysis of the Recombinant Epitope of Herpes Simplex Virus Type 2 Glycoprotein G
Xiao-Hong WANG ; Hai-Rong LU ; Gang ZHANG ; Shao-Juan CHEN ; De-Xin HUANG ; Ling-Yun LI ; Feng LIN ;
China Biotechnology 2006;0(09):-
A fragment containing amino acid residues 561~578 of HSV-2 glycoprotein G(gG2) was obtained by PCR assembling technique,and doubly cloned into vector pET-KDO.The recombinant plasmid was transformed to BL21(DE3)plysS.Fusion protein,of molecular weight about 39kDa was highly expressed by induction of IPTG.Western blot result showed the fusion protein had good antigenicity.After putification and digestion,the purity reached 95%.The digested purified protein was analysed by ELISA and showed good sensitivity and specificity.The recombinant protein should be useful for type-specific serodiagnosis of HSV-2.
7.Management of oral mucositis in patients with head and neck cancer receiving chemoradiotherapy and/or molecular targeted therapy
Xin-Xin ZHANG ; Lin MA ; Jia-Ling WANG ; Wen-Ming WU ; Lin-Chun FENG ; De-Liang HUANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(6):505-508
Objective To investigate the prevalence and treatment of oral mucositis caused by concurrent chemoradiotherapy and/or molecular targeted therapy in the patients with advanced squamous cell carcinoma of the head and neck. Methods A retrospective study of the incidence and treatment of oral mucositis was performed in 179 patients (155 male and 24 female; 124 patients at stage Ⅲ and 55 patients at stage Ⅳ) receiving concurrent chemotherapy and (or) molecular targeted therapy between November 2007 and November 2010. Grade Ⅰ , Ⅱ , Ⅲ and Ⅳ oral mucositis occurred respectively in 49, 50, 67 and 13 patients. All the patients received oral mucositis prophylaxis. After the occurrence of oral mucositis, conventional treatment of mucositis combined with quinolone antibiotics were applied. Results Of the patients, 99 patients with grade Ⅰ or Ⅱ and 4 patients with grade Ⅲ oral mucositis were effectively managed by conventional treatment; 76 patients with grade Ⅲ or Ⅳ oral mucositis were also significantly controled by conventional treatment plus antibiotics. After the treatments, all patients with oral mucositis were under control, with the decrease in the grade of oral mucositis, the reduction of oral pain and the improvement in ability to eat None of them had radiation treatment breaks. Conclusions Combined modality therapy can effectively control chemoradiation-induced oral mucositis in patients with head and neck squamous cell carcinoma, grade Ⅰ and Ⅱ oral mucositis were cured by conventional treatment and quinolone antibiotics play a key role in the treatmeats for grade Ⅲ and Ⅳ oral mucositis.
8.Molecular mechanism of effect of wenban humai granule on stability of atheromatous plaque.
Bao-ting ZHANG ; Qian-lin YAN ; De-xin YAN ; Zhi LI ; Yong-chun YU ; Guo-ping HUANG ; De-sheng TANG ; Xin YE
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(2):154-159
OBJECTIVETo explore the molecular mechanism of wenban humai granule (WHG) in stabilizing atheromatous plaque, by observing its effect on the collagen degradation and synthesis imbalance manner in the fibrous cap of the plaque.
METHODSAtherosclerosis (AS) rabbit model established by feeding high fat diet. The changes of protein and mRNA expression of macrophage CD68, metalloproteinase-1 (MMP-1), alpha-smooth muscle actin (alpha-SMA) and collagen I (C-I) in model rabbits' neo-genesic intima were determined by immunohistochemical stain and in situ hybridization methods before and after treatment as well as before and after modeling.
RESULTSAfter being fed with high fat diet for 7 weeks, the protein and mRNA expression of macrophage CD68, MMP-1 in neo-genesic intima of aorta in the model rabbits significantly increased, these changes could be significantly restored after 8 weeks treatment with WHG or simvastatin. At the same time, the expressions of alpha-SMA protein and C-I protein and mRNA slightly increased due to the immigration of SMC in aortic media to neo-genesic intima, these expressions could be further increased after WHG treatment but showed a reducing trend after simvastatin treatment (P < 0.05 and P < 0.01). In the whole course, positive correlation was shown between protein expressions of CD68 and MMP-1 (r = 0.952, P < 0.01) and also between these of alpha-SMA and C-I (r = 0.793, P < 0.01).
CONCLUSIONWHG affects the collagen degradation and synthesis imbalance in the fibrous cap of the plaque to stabilize plaque through bi-directional regulation, up-regulating synthesis thesis factors and down-regulating degradation factors, while simvastatin perform its action on plaque stability by down-regulating degradation factors alone.
Actins ; metabolism ; Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Aorta ; pathology ; Arteriosclerosis ; drug therapy ; metabolism ; pathology ; Collagen ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Macrophages ; metabolism ; Matrix Metalloproteinase 1 ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Random Allocation
9.The chest image appearances of penicilliosis marneffei in patients with AIDS
Jin-Xin LIU ; Xiao-Ping TANG ; Song-Feng JIANG ; Lie-Guang ZHANG ; Bi-Hua CHEN ; Hong-Ling SHI ; Wu-Zhi HUANG ; De-Yang HUANG ;
Chinese Journal of Radiology 2001;0(03):-
Objective To study the chest image appearances of penicilliosis marneffei(PSM)in patients with acquired immune deficiency syndrome(AIDS).Methods Chest imaging features of PSM in 36 patients with AIDS were retrospectively analyzed.Results Radiographic features of infiltrative lesions and focal lung consolidation were found in 14 cases(38.89%),in which 2 cases were with single lung disease(5.56%)and 12 cases with bilateral lung involvement(33.33%).Eight cases had diffuse lesions (22.22%),10 cases had reticular image patterns(27.78%),9 cases had nodular patterns(25.00%), 7 cases had ground-glass shadows(19.44%),6 cases had diffuse miliary lesions(16.67%),4 cases had enlarged bilar and enlarged mediastinum lymph nodes(11.11%).Cystic lesions was found in 5 cases (13.89%).Four cases had pleural effusion(11.11%),and 2 cases had nodular bump(5.56%). Pericardial effusion and pneumothorax each appeared in 1 case(2.78%).By HRCT,infiltrative lesion and focal lung consolidation were found in 32 patients(88.89%),in which 4 cases were with single lung lesions (11.11%)and 28 cases were with bilateral lung lesions(77.78%).Thirteen cases had diffuse lesions (36.11%),10 cases had pulmonary interstitial hyperplasia(27.78%),9 cases had nodular patterns (25.00%),8 cases had ground-glass shadows(22.22%),9 cases had diffuse miliary lesions(25.00%), 21 cases had enlarged lymph nodes in the mediastinum(58.33%).Cystic lesions were found in 8 cases (22.22%).Thirteen cases had pleural effusion(36.11%),and 2 cases had nodular bump(5.56%). Pericardial effusion and pneumothorax each appeared in 1 case(2.78%).Conclusion The image appearances of PSM in patients with AIDS include infiltrative lesions or focal lung consolidation,ground- glass shadow,enlarged hilar and mediastinum lymph nodes,pleural effusion,interstitial involvement or reticular image pattern(pulmonary interstitial hyperplasia),diffuse miliary lesion,and cystic lesion.
10.Anticoagulation therapy in Chinese patients with non-valvular atrial fibrillation: a prospective, multi-center, randomized, controlled study.
Ke-ping CHEN ; Cong-xin HUANG ; De-jia HUANG ; Ke-jiang CAO ; Chang-sheng MA ; Fang-zheng WANG ; Shu ZHANG
Chinese Medical Journal 2012;125(24):4355-4360
BACKGROUNDNon-valvular atrial fibrillation is associated with an increased risk of ischemic stroke; however, the appropriate intensity of anticoagulation therapy for Chinese patients has not been determined. The purpose of this study was to compare the safety and the efficacy of standard-intensity warfarin therapy, low-intensity warfarin therapy, and aspirin therapy for the prevention of ischemic events in Chinese patients with non-valvular atrial fibrillation (NVAF).
METHODSA total of 786 patients from 75 Chinese hospitals were enrolled in this study and randomized into three therapy groups: standard-intensity warfarin (international normalized ratio (INR) 2.1 to 2.5) group, low-intensity warfarin (INR 1.6 to 2.0) group and aspirin (200 mg per day) group. All patients were evaluated by physicians at 1, 3, 6, 9, 12, 15, 18, 21 and 24 months after randomization to obtain a patient questionnaire, physical examination and related laboratory tests.
RESULTSThe annual event rates of ischemic stroke, transient ischemic attack (TIA) or systemic thromboembolism were 2.6%, 3.1% and 6.9% in the standard-intensity warfarin, low-intensity warfarin and aspirin groups, respectively (P = 0.027). Thromboembolic event rates in both warfarin groups were significantly lower than that in the aspirin group (P = 0.018, P = 0.044), and there was no significant difference between the two warfarin groups. Severe hemorrhagic events occurred in 15 patients, 7 (2.6%) in the standard-intensity warfarin group, 7 (2.4%) in the low-intensity warfarin group and 1 (0.4%) in the aspirin group. The severe hemorrhagic event rates in the warfarin groups were higher than that in the aspirin group, but the difference did not reach statistical significance (P = 0.101). The mild hemorrhagic and total hemorrhagic event rates in the warfarin groups (whether in the standard-intensity warfarin group or low-intensity warfarin group) were much higher than that in the aspirin group with the annual event rates of total hemorrhages of 10.2%, 7.6% and 2.2%, respectively, in the 3 groups (P = 0.001). Furthermore, there was no significant difference in all cause mortality among the three study groups.
CONCLUSIONIn Chinese patients with NVAF, the warfarin therapy (INR 1.6 - 2.5) for the prevention of thromboembolic events was superior to aspirin.
Aged ; Aged, 80 and over ; Anticoagulants ; administration & dosage ; therapeutic use ; Aspirin ; administration & dosage ; therapeutic use ; Atrial Fibrillation ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Warfarin ; administration & dosage ; therapeutic use