2.High-level Secretion Expression of Human ScFv Against Botulinum Neurotoxin A in Pichia pastoris*
Hui WANG ; Jun YIN ; Xiao-Jun HOU ; Hong-Guang XING ;
Microbiology 1992;0(02):-
The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72~84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.
3.Embryonic mouse pancreas transplantation for treatment of diabetes mellitus
Aijing SHAN ; Jun YANG ; Xi CHEN ; Guang NING ; Zhengming WANG
Chinese Journal of Tissue Engineering Research 2011;15(44):8237-8242
BACKGROUND: Embryonic pancreatic tissue is characterized by its abundance, potent in proliferation & differentiation, and minimal immunological rejection. It is widely considered as potential pancreatic endocrinological stem cells resource for treating diabetes mellitus.OBJECTIVE: To investigate the embryonic mouse pancreatic tissue isolation technique and observe the recipients' blood glucose regulatory effects of the grafted embryonic pancreas in an experimental diabetes mellitus mouse model.METHODS: Pancreatic tissue from C57B1/6 mouse embryos at embryonic days 11.5-16.5 was isolated under the stereomicroscope. C57BL/6 mouse models of streptozocin-induced diabetes mellitus were established and then randomly divided into two groups: transplantation group, in which, five pieces of pancreatic tissue of mice at embryonic 16.5 days were transplanted into mouse renal capsule, and sham-operated control group, in which, 0.05 mL RPMI1640 culture medium was injected into mouse renal capsule. When blood glucose level of the transplantation group mouse was≤ 11.2 mmol/L, the endocrine function of embryonic pancreatic tissue transplanted was detected by IPGTT and IPITT methods and then the transplanted graft was removed for observing the blood glucose relapse.RESULTS AND CONCLUSION: Nearly intact pancreatic tissue of mice at embryonic days 11.5-16.5 could be isolated through the use of stereomicroscope. Pancreatic tissue morphology and color of mice ≤ embryonic 12.5 days were difficultly distinguished from adjacent tissue and they could only be isolated carefully according to the relationship with adjacent organs. Pancreatic tissue of mice > embryonic 12.5 days exhibited initial endocrinological tissue morphology mimic white cauliflower. Histological and ELISA examinations showed that embryonic pancreatic tissue could express and secrete insulin and the insulin level was gradually increased with developmental time. Embryonic pancreatic tissue could grow beneath the recipient renal capsule. The insulin and glucagon expression in the post-transplantational pancreatic tissue graft was increased compared with prior to transplantation. These results suggest that pancreatic tissue is a potential stem cell resource for treating the diabetes mellitus.
4.APPLICATION OF CELL ENZYME IMMUNOASSAY FOR DETECTION AND ANALYSIS OF MONOCLONAL ANTIBODIES TO HUMAN COLORECTAL ADENOCARCINOMA
Jun WANG ; Caixia LIN ; Hui YU ; Guang LI
Chinese Journal of Immunology 1985;0(02):-
A solid-phase cell enzyme-linked immunoassay is described for screeningand analyzing monoclonal antibodies against the cell surface antigensof human colorectal adenocarcinoma.Horse-radish peroxidase conjugatedProtein A was used to detect the binding of mouse monoclonal antibodiesto human colorectal adenocarcinoma.HR_(8348) cells which had been cultured inand then fixed to the wells of microtiter plates by using glutaraldehyde.Thismethod was found to be specific,reproducible and practical,and especiallyto be advantageous for the large scale screening and analyzing of monoclo-nal antibodies with a panel of cell types.
5.Preoperative diagnosis and treatment of primary non-specific ureteritis(report of 3 cases)
Hong-Yu ZHUANG ; Yong-Guang JIANG ; Jun-Sheng WANG ;
Chinese Journal of Urology 2001;0(10):-
Objective To evaluate the preoperative diagnosis and treatment of primary non-specific ureteritis,and to better understand this disease.Methods Three cases of primary non-specific ureteritis (1 man and 2 women)were reported.Their age was 28,38,68 years,respectively.One ease was found to have hydronephrosis on B-ultrasound at physical examination with no symptoms,and 2 had abdominal pain. The lesions were detected in lower part of the ureter in 2 cases,and in middle in I.The patients were diag- nosed preoperatively with combined use of MRU and ureteroscopy.Partial resection of the diseased part of the ureter was performed in all the cases.Results Pathological findings confirmed the diagnosis of non-spe- cific ureteritis.Microscopy showed necrotic tissue and inflammatory cell infiltration of ureteral mucosa,dilated and congested blood vessels,hypertrophic muscular layer,and proliferation of fibrous tissue.During the fol- low-up of 2 years,the patients had no symptoms and were free from hydronephrosis.Conclusions The eti- ology of primary non-specific ureteritis is unclear,and no typical symptoms can be found clinically.Combined use of MRU and ureteroscopy is helpful for definite diagnosis of the disease.Partial resection of the diseased part of the ureter has good results.
6.Analysis of the direction and degree of static cyclotorsion component and dynamic cyclotorsion component in corneal refractive surgery
Bei, HUANG ; Dan-Dan, ZHAO ; Guang-Hua, ZHAO ; Jun, WANG
International Eye Science 2016;16(7):1258-1260
AIM: To analyze the direction and degree of static cyclotorsion component (SCC) and dynamic cyclotorsion component (DCC) in corneal refractive surgery.
METHODS:Retrospective analysis. Totally 130 patients (260 eyes) with corneal refractive surgery in our hospital, according to the operation method were divided into femtosecond laser - assisted laser in situ keratomileusis (FS-LASIK) group and T-photorefractive keratectomy (T-PRK) group, the differences of the parameters of the two groups were compared; the differences of SCC success rate, SCC, DCC, and the eyeball rotation direction were compared between the two groups; correlation analysis on SCC, DCC and the parameters of postoperative patients were performed.
RESULTS: High order aberrations and spherical aberration in the T-PRK group after operation was higher than those of FS - LASIK group, and the difference was statistically significant (P<0. 05); in T-PRK group SCC in the operation was successful in 98 eyes, the success rate was 81. 7%; in FS-LASIK group SCC in the operation was successful in 82 eyes, the success rate is 58. 6%, the difference of SCC success rate between the two groups was statistically significant ( P < 0. 05 ); SCC in T - PRK group was 3. 52o ±2. 17o and FS-LASIK group was 3. 49o ±2. 26o, there was no significant difference (P>0. 05); DCC in T-PRK group (2. 86o±1. 14o) was higher than that of FS-LASIK group ( 2. 17o ± 1. 09o), and the difference was statistically significant (P<0. 05). There was no statistical difference in the direction of rotation of the eyeball in operation between the two groups (P>0. 05). The SCC of subjects in operation was positively correlated with UCVA, BCVA, spherical equivalent refraction and high order aberrations ( P < 0. 05 ); the DCC of subjects in operation was positively correlated with UCVA and high order aberrations (P<0. 05).
CONCLUSION: The success rate of SCC in T - PRK surgery is higher than that in LASIK, DCC in T - PRK surgery is higher than that in LASIK, and accurate measurement of SCC and DCC can be effective to compensate for it.
7.Significance of Computed Tomography in Diagnosis of Bronchial Foreign Body in Children
yan, SUN ; shao-hua, WANG ; rong-jun, LIN ; guang-feng, JIANG ; ying, TIAN ; zhi-jun, CHEN
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate clinical significance of computed tomography (CT) scan in diagnosis of bronchial foreign body in children.Methods Twenty-one suspected children with bronchial foreign body were studied with spiral CT cross-section scan and coronal reconstruction and diagnosis was confirmed with bronchoscopy.Results The foreign body was displayed in all of 21 cases. CT scan showed foreign body was located in right main bronchial 12 cases, right middle bronchial 1 case, right inferior lobar bronchial 2 cases and left main bronchial 6 cases. Foreign bodies were extracted with bronchoscopy.Conclusion CT scan can display and locate accurately foreign body in bronchial of children,and has very important diagnostic value in patients having atypical histories, clinical and radiological findings.
8.Correlation between p53 gene mutations and p53 protein overexpression in esophageal squamous cell carcinoma.
Guang LI ; Zhao-xia LIU ; Jing HUANG ; Jun-mei WANG ; Quan-hong WANG
Chinese Journal of Pathology 2005;34(12):802-804
Adult
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Aged
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Carcinoma, Squamous Cell
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genetics
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metabolism
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Esophageal Neoplasms
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genetics
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metabolism
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Female
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Gene Expression Regulation, Neoplastic
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Genes, p53
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Humans
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Loss of Heterozygosity
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Male
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Middle Aged
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Point Mutation
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Tumor Suppressor Protein p53
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genetics
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metabolism
9.Investigation of PML-RAR? fusion gene in acute promyelocytic leukemia by real-time quantitative reverse transcription PCR
Hong YUAN ; Nan WANG ; Shi-Jun LI ; Jing WANG ; Hong ZHU ; Xiao-Guang XIAO ; Min HUANG ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To investigate the relapse risk assessment for patients with acute promyelocytic leukemia(APL)through testing PML-RAR? fusion gene by real-time quantitative polymerase chain reaction(RQ-PCR).Methods Relative copies of PML-RAR? fusion gene were measured in 25 patients with APL in phases of first diagnosis,complete remission(CR)and relapse.The minimal residual disease(MRD)situations were also monitored in 6 of the 25 patients.Results Different PML-RAR? fusion gene expression levels were observed in patient groups of different phases of the disease.(P
10.Chinmedomics: a new strategy for research of traditional Chinese medicine.
Ai-hua ZHANG ; Hui SUN ; Guang-li YAN ; Ping WANG ; Ying HAN ; Xi-jun WANG
China Journal of Chinese Materia Medica 2015;40(4):569-576
Syndrome and formulae (or prescription) are two key issues in traditional Chinese medicine (TCM) and the premise research for material basis of TCM. However, vagueness of syndromes and complexity of formulae greatly limited the evaluation to syndromes and effective substance basis of prescription. Therefore, how to solve the evaluation of syndromes, confirming the efficacy material basis in prescription are the current hot issues of international concern. To solve these problems, establishing chinmedomics by integrated serum pharmacochemistry of TCM with metabolomics technology, that is a unique method of TCM research, made outstanding contributions in solving international concerns such as the effectiveness and security aspects of TCM. On the basis of the biological characterization of syndrome, the metabolic profiling of animal models of TCM syndrome, and related metabolic fingerprints as well as metabolic biomarkers were established to evaluate the overall effects of TCM formulae and corresponding relationship of syndrome-formulae. The active constituents were screened using the plotting of correlation between (endogenous) marker metabolites and (exogenous) serum constituents (PCMS), and is ongoing verification by further biological experiments. Correlation analysis between the ingredients in the body after oral formulae and endogenous markers in vivo can be used to clarify the active ingredients and synergistic properties. This method was successfully applied for rapid discovery of potentially bioactive components and metabolites from TCM, and through a series of studies on the chinmedomics, it proved that the established method could help to explore the effective substance for further research of TCM. As a new research approach, Chinmedomics is the best method to fit the holistic concept of TCM, and it can not only interpret the essence of syndrome but also elucidate the scientific connotation of Chinese medical formulae.
Animals
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Diagnosis, Differential
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Drug Prescriptions
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Drug Therapy
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Drugs, Chinese Herbal
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analysis
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pharmacokinetics
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Humans
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Medicine, Chinese Traditional