3.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.
4.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.
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.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
8.Total Hip Arthroplasty Using Modular Trabecular Metal Acetabular Components for Failed Treatment of Acetabular Fractures: A Mid-term Follow-up Study.
De-Yong HUANG ; Liang ZHANG ; Yi-Xin ZHOU ; Chun-Yu ZHANG ; Hui XU ; Yong HUANG
Chinese Medical Journal 2016;129(8):903-908
BACKGROUNDPorous-coated cups have been widely used in acetabular reconstruction after failed treatment of acetabular fractures, and good results have been reported with the use of these cups; however, the durability and functionality of modular trabecular metal (TM) acetabular components in acetabular reconstruction after failed treatment of acetabular fractures remain unclear. This study aimed to examine the radiographic and clinical outcomes associated with the use of modular TM acetabular components for failed treatment of acetabular fractures to assess the durability and functionality of these components in acetabular reconstruction after failed treatment of acetabular fractures.
METHODSA total of 41 patients (41 hips) underwent total hip arthroplasty (THA) using modular TM acetabular components for failed treatment of acetabular fractures at our hospital between January 2007 and December 2012. Among these patients, two were lost to follow-up. Therefore, 39 patients (39 hips) were finally included in this study. The Harris hip score before and after the surgery, satisfaction level of the patients, and radiographic results were assessed.
RESULTSThe mean Harris hip score increased from 34 (range, 8-52) before surgery to 91 (range, 22-100) at the latest follow-up examination (P < 0.001). The results were excellent for 28 hips, good for six, fair for three, and poor for two. Among the 39 patients, 25 (64%) and 10 (26%) were very satisfied and somewhat satisfied, respectively. All cups were found to be fully incorporated, and no evidence of cup migration or periacetabular osteolysis was noted.
CONCLUSIONSDespite the technically demanding nature of the procedure, THA using modular TM acetabular components showed good durability and functionality and may be an effective reconstruction option for failed treatment of acetabular fractures.
Acetabulum ; diagnostic imaging ; injuries ; surgery ; Adult ; Aged ; Arthroplasty, Replacement, Hip ; adverse effects ; Female ; Follow-Up Studies ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Treatment Failure
9.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
10.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.