2.Construction of Salmonella Pullorum C79-13ΔcrpΔasd mutant balanced-lethal host-vector system and its biological features
Songbiao CHEN ; Jing LI ; Ke SHANG ; Chuan YU ; Chunjie ZHANG ; Xiangchao CHENG ; Yinju LI ; Zhanqin ZHAO
Chinese Journal of Immunology 2014;(8):1083-1087,1092
In order to develop a safer vaccine strain exploit Salmonella Pullorum vaccine strain as a live vaccine vector.Methods:AΔcrpΔasd mutant of S.pullorum C79-13 strain was constructed and it was developed E.coli donor strain mutant was generated through the two-step method introduced by χ7213 ( pREΔasd) was conjugated with the recipient of C 79-13Δcrp.The C79-13ΔcrpΔasd the transduction of recombinant suicide plasmid .Results:PCR and sequencing results showed that ΔsipBSL1344 was suc-cessfully constructed.The further study indicated that foreign DAP must be supplied for the ΔcrpΔasd mutant to grow,in addition,the asd gene was transmitted stably .Compared with C79-13 strain,the O antigens was identical to C79-13 strain,but the growth velocity was reduced significantly ,significantly reduced virulence .Conclusion: The ΔcrpΔasd mutant can accept asd+plasmid to construct host-vector balance lethal system , and this system can be used to express foreign gene efficiently and to develop potential oral multivalent vaccines.
3.Construction and characterization of host-vector balanced lethal system of attenuated salmonella choleraesuisΔcrpΔcyaΔasdC78-1 (pYA3493)
Ke SHANG ; Junfeng ZHANG ; Xiangchao CHENG ; Chunjie ZHANG ; Yinju LI ; Guihua CHEN ; Yunfei YAN ; Chongkai ZHAI ; Zhanqin ZHAO
Chinese Journal of Immunology 2015;(3):358-363
Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.
4.Effect of low-frequency repetitive transcranial magnetic stimulation promotes recovery of cognitive function and functional independence in patients with traumatic brain injury
Ju-Ke HUANG ; Rong-Kang MAI ; Shang-Jie CHEN
Chinese Journal of Neuromedicine 2009;8(8):854-856
Objective To observe the effect of low-frequency repetitive transcranial magnetic stimulation in improving the memory function and quality of life of patients with traumatic brain injury. Methods Sixty patients with brain injury were randomly divided into the treatmem group and control group. The patients in the control group received conventional treatments (including medications, acupuncture and exercise training), and those in the treatment group were given additional low-frequency repetitive transcranial magnetic stimulation treatment. Scores of the Rivermead Behavioral Memory Test (RBMT) and Functional independence Measure (FIM) were recorded before and after the treatments and compared between the two groups. Results The patients in both of the groups showed significant improvements in RBMT and FIM scores after the treatments (P<0.05). The improvements in RBMT and FIM scores after the treatments differed significantly between the two groups (P<0.05). Conclusion Low-frequency repetitive transcranial magnetic stimulation can significantly improve the cognitive function and functional independence of patients with traumatic brain injury.
5.Expression of chemokine receptor-7 in oral squamous cell carcinoma and its relation to cervical lymph node metastasis
Zheng-Jun SHANG ; Ke LIU ; Xin-Ming CHEN
Chinese Journal of Stomatology 2008;43(10):592-596
Objective To investigate the expression of chemokine receptor CCR7 in oral squamous cell carcinoma (OSCC) and its possible role in directional neck lymph node metastasis. Methods The expression of chemokine receptor CCR7 was examined in 85 cases of OSCC,normal oral mucosa,and TcaS113 and adenoid cystic carcinoma (ACC-M) cell lines using immunohistochemistry,RT-PCR and Western blotting.The relationship between the expression and clinicopathological factors of OSCC was analyzed.After treatment with or without CCR7 antibody,in vitro adhesion assay was performed to compare the adhesion of Tea8113 and ACC-M cells to lymph nodes. Results CCR7 was expressed in 66% (56/85)of OSCC cases,and there was a significant difference in CCR7 expression between metastasis group (31/39,79%) and non-metastasis group (25/46,54%).RT-PCR and Western blotting also confirmed the presence of positive CCR7 expression in OSCC. Tea8113 cells expressing CCR7 showed stronger ability to adhere to lymph nodes as compared to CCR7-negative ACC-M cells and could be actively inhibited by CCR7 antibody.Conclusions CCR7 may play an important role in the development and lymph node metastasis of oral squamous cell carcinoma.
6.Long-term follow-up study of dilation with temporary cardia stent in different diameters for achalasia
Ying-Sheng CHENG ; Ming-Hua LI ; Ren-Jie YANG ; Wei-Xiong CHEN ; Ni-Wei CHEN ; Qi-Xing ZHUANG ; Ke-Zhong SHANG ;
Journal of Interventional Radiology 2006;0(11):-
Objective To determine the curative effect of dilation for achalasia with temporary cardia stent in different diameters based on a long-term follow-up.Methods The study cohort was comprised of 135 patients of achalasia.Among them differentiated by stent diameters as followings:30 patients were treated under fluoroscopy with dilation of temporary cardia stent in 20 mm diameter(group A), 30 patients with dilation of temporary eardia stent in 25 mm diameter(group B),and 75 patients with dilation of temporary cardia stent in 30 mm diameter(group C).135 cardia stents were temporarily placed in the 135 patients and withdrawn after 3 -5 days via gastroscopy.All the stents were inserted and withdrawn successfully.The follow-up in all groups lasted 6-128 months.Results Six(20.0%)out of 30 patients,6(20.0%)out of 30 patients,5(22.7%)out of 22 patieuts,6(37.5%)out of 16 patients,5 out of 9 patients,3 out of 3 patients in group A exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively. Four(13.3%)out of 30 patients,4(13.3%)out of 30 patients,3(13.0%)out of 23 patients,4(22.2%)out of 18 patients,5(45.5%)out of 11 patients,and 3 out of 4 patients in group B exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.No(0.0%)out of 75 patients, 1(1.5%)out of 66 patients,4(8.3%)out of 48 patients,6(18.2%)out of 33 patients,6(33.3%)out of 18 patients,2 out of 5 patients in group C exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.Conclusion Dilation with temporary cardia metal stent in 30 mm diameter is the best dilation for achalasia in long-term follow-up.
7.The binding of MBL to common bacteria in infectious diseases of children.
Shi-qiang SHANG ; Guo-xian CHEN ; Jie SHEN ; Xiao-hong YU ; Ke-Yi WANG
Journal of Zhejiang University. Science. B 2005;6(1):53-56
OBJECTIVETo purify Mannan-binding lectin (MBL) from human serum and detect its binding ability to several kinds of bacteria common in infectious diseases of children.
METHODSMBL was purified from human serum by affinity chromatography on mannan-Sepharose 4B column. Its binding ability to eight species, 97 strains of bacteria was detected by enzyme-linked lectin assay (ELLA).
RESULTSMBL has different binding ability to bacteria and shows strong binding ability to Klebsiella ornithinolytica and Escherichia coli, but shows relatively lower binding ability to Staphylococcus haemolyticus, Enterobacter cloacae and Staphylococcus epidermidis. To different isolates of Klebsiella pneumoniae, Haemophilus influenzae and Staphylococcus aureus, MBL shows quite different binding ability.
CONCLUSIONSMBL has different binding ability to different bacteria, and has relatively stronger binding ability to Gram-negative bacteria. Its binding ability to different isolates of certain kinds of bacteria is quite different.
Bacteria ; classification ; metabolism ; Child ; Child, Preschool ; Communicable Diseases ; microbiology ; Humans ; Mannose-Binding Lectin ; blood ; metabolism ; Protein Binding ; Species Specificity
8.A multi-center randomized double-blind placebo-controlled trial of Xiongshao Capsule in preventing restenosis after percutaneous coronary intervention: a subgroup analysis of senile patients.
Qing-hua SHANG ; Hao XU ; Xiao-yan LU ; Chuan WEN ; Da-zhuo SHI ; Ke-ji CHEN
Chinese journal of integrative medicine 2011;17(9):669-674
OBJECTIVETo evaluate the safety and efficacy of Xiongshao Capsule (XS), consisting of Chuangxiongol and paeoniflorin, in preventing restenosis after percutaneous coronary intervention (PCI) in senile coronary heart disease (CHD) patients.
METHODSA multi-center, randomized, double-blind, placebo-controlled trial was conducted. A total of 335 CHD patients were randomly assigned to treatment with oral administration of XS, or a placebo for 6 months after successful PCI. A clinical follow-up was performed at 1, 3 and 6 months after PCI and an angiographic follow-up was scheduled at 6 months. The primary endpoint was angiographic restenosis defined as a luminal stenosis ≥ 50% in follow-up. The secondary endpoints were combined incidence of death, target lesion nonfatal myocardial infarction, repeat target-vessel angioplasty, and coronary artery bypass graft surgery (CABG). The follow-up for the above clinical endpoint events was continued to 1 year after PCI.
RESULTSThe subgroup analysis of 152 senile patients (68 cases angiographic follow-up) showed that the restenosis rates tended to reduce in the XS group as compared with that in the placebo group (24.32% vs. 38.71%, P > 0.05), and the minimum lumen diameter (MLD) significantly increased in the follow-up (2.15 ± 0.84 for XS vs. 1.73 ± 0.91 for placebo, P < 0.05). The incidence of recurrent angina at 3 and 6 months after PCI was also significantly reduced in the XS group (4.11% and 12.33%) as compared with those in the placebo group (17.72% and 43.04%), but there was no significant difference in the combined incidence of clinical outcomes (6.85% in the XS group vs. 11.39% in the placebo group, P > 0.05). No significant adverse reactions occurred within the 6-month follow-up period in the XS group.
CONCLUSIONAdministration of XS in addition to standardized Western medication for 6 months is demonstrated to be safe and effective in reducing post-PCI recurrent angina and inhibiting luminal restenosis after PCI in senile CHD patients.
Aged ; Angina Pectoris ; complications ; diagnostic imaging ; epidemiology ; Angioplasty, Balloon, Coronary ; adverse effects ; Capsules ; China ; epidemiology ; Coronary Angiography ; Coronary Restenosis ; diagnostic imaging ; drug therapy ; etiology ; prevention & control ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Endpoint Determination ; Female ; Humans ; Male ; Placebos ; Recurrence
9.Clinical curative effect observation of double tube method in the treatment of esophagojejunostomy leakage after laparoscopic for total gastrectomy.
Xiang GENG ; Hai Liang LI ; Chen Yang GUO ; Hong Tao HU ; Hong Tao CHENG ; Quan Jun YAO ; Chuang SHANG ; Ke ZHAO
Chinese Journal of Gastrointestinal Surgery 2022;25(7):627-631
10.Effects of Panax quinquefolium saponin on phosphatidylinositol 3-kinase/serine threonine kinase pathway of neonatal rat myocardial cells subjected to hypoxia.
Chun-yu GUO ; Xiao-juan MA ; Jing-shang WANG ; Ying SHI ; Xin LIU ; Hui-jun YIN ; Ke-ji CHEN
Chinese journal of integrative medicine 2015;21(5):384-388
OBJECTIVETo explore the effects of Panax Quinquefolium Saponin (PQS) on phosphatidylinositol 3-kinase/serine threonine kinase (PI3K/Akt) pathway of neonatal rat myocardial cells subjected to hypoxia.
METHODSNeonatal rat myocardial cells were cultured in vitro. After the myocardial cell injury was induced by hypoxia, the cells were randomized into 5 groups: the normal group, the model group, the positive control group (Ciclosporin A, 2 µ mol/L), the low-dose PQS group (PQSL, 25mg/L), and the high-dose PQS group (PQSH, 50 mg/L). Morphology and behavior of myocardial cells were observed under an inverted microscope. Apoptosis rate and lactate dehydrogenase (LDH) leakage rate of myocardial cells were determined by colorimetry. Mitochondrial transmembrane potential was assessed using a fluorexon laser. Phospho-glycogen synthase kinase (GSK)-3β and phospho-Akt as well as cytochrome C were determined by Western blot
RESULTSLDH leakage in the Ciclosporin A group, PQSH group and PQSL group reduced progressively compared with the model group (P<0.05). Akt and GSK-3β was strongly phosphorylated after treatment with Ciclosporin A and PQS compared with the model group (P<0.05, P<0.01). Compared with the model group (16.41±1.74; 35.28±6.30), both the integrated optical density of mitochondrial permeability transition pore (MPTP) and the mitochondrial transmembrane potential significantly increased in the PQSH group (42.74±2.12; 71.36±6.54) and the PQSL group (39.58±1.49; 66.99±5.45; P<0.05, P<0.01). However, the protein of cytochrome C outside the mitochondrion decreased in the PQSH group (273.66±14.61) and the PQSL group (259.62±17.31) compared with the model group (502.41±17.76; P<0.05).
CONCLUSIONThrough activation of the PI3K/Akt pathway and inhibition of the MPTP, PQS might protect the heart against ischemia injury and apoptosis of myocardial cells.
Animals ; Animals, Newborn ; Cell Hypoxia ; drug effects ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; L-Lactate Dehydrogenase ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; metabolism ; Myocytes, Cardiac ; cytology ; drug effects ; enzymology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; drug effects ; Protein-Serine-Threonine Kinases ; metabolism ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; Signal Transduction ; drug effects