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.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.
5.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.
6.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.
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.Effect of Melatonin on CD4~+CD_(25)~+ Regulatory T Cell and Airway Inflammation in Asthmatic Rat
min, WANG ; jun-lan, YAO ; an-xiu, XIONG ; guang-huan, ZHANG ; qun-xing, WANG ; chun-hua, LUO
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To explore the effect of Melatonin(MT) on CD4+CD25+ regulatory T cell (CD4+CD25+Tr)and airway inflammation in asthmatic rat.Methods Thirty-two SD rats were randomly divided into 4 groups,8 rats in each group.Asthmatic group:rats were immunized on day 1 and 7 by intraperitoneal inject of mixture of ovalbumin(OVA) and aluminumhydroxide.From day 14,the animals were allenged with aerosolized OVA for 20 min per day for 7 consecutive days.MT group:OVA-sensitized rats were injected intraperitoneally with 0.1 mg/kg MT 30 min before each OVA challenge.Dexamethasone group:OVA-sensitized rats were injected intraperitoneally with 0.5 mg/kg Dexamethasone 30 min before each OVA challenge.Control group:OVA for inhalation and MT for intraperitoneal injection was replaced with saline.After the last challenge,peripheral blood was stained to count the percentage of eosinophil(EOS).Then the rats were lavaged and total leukocytes counts in bronchoalveolar lavage fluid(BALF) were performed after staining with Wright-Giemsa staining.The EOS counts around the airway was counted after the histological section of lung staining with hematoxylin and eosin staining.The serum level of immunoglobulin E(IgE) was detected by immunoenhancement.The change of CD4+CD25+Tr was assessed with flow cytometry.SPSS 10.0 software was applied to analyze data. Results In asthmatic rats,the CD4+CD25+ Tr/ CD4+T cells ratio had significant negative relationship with the EOS counts around the airway and the total leukocytes counts in BALF (r=-0.73 P0.05).There was a significant decrease in the percentage of the eosinophils in peripheral blood,the eosinophil counts around the airway,the total leukocytes counts in BALF and the serum level of IgE in MT group compared with asthmatic group (Pa
9.Genetic polymorphism of T6235C mutation in 3' non-coding region of CYP1A1 and GSTM1 genes and lung cancer susceptibility in the Mongolian population
Fuhou CHANG ; Minjie WANG ; Jun QI ; Qin YIN ; Lei FAN ; Ruilan HAN ; Guang WANG
Journal of Pharmaceutical Analysis 2009;21(4):225-229
Objective To estimate the relative risk for lung cancer associated with genetic polymorphism of T6235C mutation in 3' non-coding region (Msp Ⅰ) of cytochrome P450 1A1 (CYP1A1) and glntathione S-transferase M1 (GSTM1) in the Mongolian population in Inner Mongolian Region of China. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR methods were used to analyze blood samples obtained from 263 case subjects and 263 control subjects to determine their genotypes for CYP1A1 and GSTM1.Control subjects were matched with case subjects by ethnic background, age and gender. Results The frequencies of the variant CYP1A1 genotypes (CYP1A1C) and GSTM1-null in lung cancer groups were higher than those in control groups (38.4% vs. 28. 5% and 57.8% vs. 48.0%). The individuals who corried with CYP1A1C genotype had a significantly higher risk of lung cancer (OR=1.56, 95% CI=1.08 to 2.25, P=0.016) than those who carried with non-variation CYP1A1 genotype. The ones who carried with GSTM1-null genotype also had a significantly higher risk of lung cancer (OR=1.49, 95% CI=1.06 to 2.10, P=0.023) than these who carried with GSTM1-present genotype.When combination of polymorphisms of CYP1A1 and GSTM1 genotypes was analyzed, the risk of lung cancer for combination of CYP1A1C and GSTM1-null genotypes was increased significantly (OR=2.084, 95e CI=1.27 to 3.42, P=0.003). Susceptibility to lung cancer was related to smoking (OR=2.10, 95% CI=1.48 to 2.98, P=0.000). Considering smoking status, the risk of lung cancer for combination of smoking and CYP1A1C genotype was remarkably increased (OR=2.76, 950/0 CI=1.74 to 4. 37, P=0.000). It was the same case with combination of smoking and GSTM1-null genotype (OR=4. 38, 95% CI=2.35 to 8.15, P=0.000). Conclusion The polymorphisms of CYP1A1C genotype and GSTM1-null are the risk factors of lung cancer in the Mongolian population in Inner Mongolia Region of China. Smoking is also related to susceptibility to lung cancer. There may be a synergetic interaction between CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer. Smoking may have a synergetic interaction with CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer.
10.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