Objective To investigate the clinical effects of the Viscoat viscoelastic combined with soft corneal contact lens for central corneal perforation.Methods Six cases were collected and treated with corneal local debridement of which diameter were less than 2.0 mm.Six cases received Viscoat viscoelastic injection into their anterior chamber.And then soft corneal contact lens were worn.The curative effect indicators such as patients' symptom,visual acuity,slit lamp examination,intraocular pressure,confocal microscope and corneal endothelial cell counts were recorded in the follow-up periods.Results All the cases were healed with the recovery time of 1 month to 2 months;After treatment,the best corrected visual acuity of patients were increased to 0.6-0.8 and average corneal endothelial cell count was (3415.5 ±279.5)mm-2.No obvious scar was left in the cornea and no serious complicatious occurred during treatment.Conclusion For traumatic corneal central perforation with diameter is 2.0 mm or less can be treated with Viscoat viscoelastic combined with soft corneal contact lens.This therapy is worthy of popularize since it's satisfied prognosis and less economic burden.

OBJECTIVETo prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.

METHODSTIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs.

RESULTSThe prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein.

CONCLUSIONThe TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.

Animals ; Antibodies, Monoclonal ; immunology ; Cloning, Molecular ; Humans ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology ; isolation & purification ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; immunology

Complete mannanase gene with two introns was cloned from Trichoderrna reesei by PCR. The two introns were then removed by overlap extension PCR. The gene encoding the mature mannanase protein was inserted into the expression vector pPIC9K, downstream of a alpha-factor signal peptide sequence. The resultant recombinant vector was named pM242. After linearized with Sac I , pM242 was transformed to Pichia pastoris GS115 by electroporation. After screening, the recombinant strain Gpmf25 that expresses the secretory protein at high level was obtained. The activity of the recombinant mannanase reached 12.5 IU/mL. Optimum pH and temperature for the recombinant enzyme were 5.0 and 80 degrees C, respectively. The enzyme was stable at pH 5.0-6.0 and maintained over 50% of original activity after incubation at 70 degrees C for 30 min.
Fungal Proteins ; biosynthesis ; genetics ; Hydrogen-Ion Concentration ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Temperature ; Trichoderma ; enzymology ; genetics ; beta-Mannosidase ; biosynthesis ; genetics

In order to explore what role relative cytokines play in acute GVHD (aGVHD) of mice transplanted with H-2 haploidentical nonmyeloablative bone marrow, a murine model was established by using C57BL/6J mouse as donor and (C57BL/6J x BALB/C) F1 mouse as the recipient. The recipient mice were given CsA and mycophenolate mofetile (MMF) for prophylaxis of aGVHD. The expression levels of IL-2, INFgamma, IL-4 and IL-10 in the spleen were detected by semi-quantitative RT-PCR at different time points after transplantation. The results showed that the expression levels of these cytokines increased slightly in the group only received nonmyeloablative conditioning. The expression levels of IL-2 and INF-gamma elevated significantly after transplantation in group 2 (without aGVHD prophylaxis), peaked at day 21 and day 14 respectively, then dropped rapidly, returned to the basal levels at day 35. The study on kinetic pattern of expression of IL-2 and INF-gamma in group 3 (with aGVHD prophylaxis) gave a similar result to group 2, but the peak value of cytokine expression in group 3 was much lower than that in group 2. The expression of IL-4 in Group 2 and group 3 all peaked at day 14, but the peak value in group 3 was higher than that in group 2, and decreased slowly in group 3. The expression of IL-10 increased gradually after transplantation, peaked at day 14, decreased after day 21, elevated again until day 35. It is concluded that the expression levels of these cytokines in the spleen all increase after H-2 haploidentical nonmyeloablative bone marrow transplantation. CsA and MMF can reduce the incidence and severity of aGVHD by down-regulating the expression levels of IL-2 and INF-gamma.
Animals ; Bone Marrow Transplantation ; immunology ; methods ; Cyclosporine ; administration & dosage ; Cytokines ; genetics ; Gene Expression ; drug effects ; Graft vs Host Disease ; prevention & control ; H-2 Antigens ; genetics ; Interferon-gamma ; genetics ; Interleukin-2 ; genetics ; Kinetics ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.

METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.

RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.

CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.

Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

The beta-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1.794. The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression P-glucosidase activity reached to 24.7 IU/mL. The optimum temperature and optimum pH of the recombinant expression P-glucosidase in BL1979 were 37 degrees C and 7.0 respectively,the purity can reach to 92.7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis, we found the fusion protein exists in dimmer, tetramer,hexamer and octamer, they all have hydrolase activity.
Bacillus ; enzymology ; Escherichia coli ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; isolation & purification ; beta-Glucosidase ; genetics ; isolation & purification ; metabolism

Bartter Syndrome ; complications ; diagnosis ; pathology ; Child, Preschool ; Humans ; Male ; Nephrosis, Lipoid ; complications ; diagnosis ; pathology ; Treatment Outcome

Humans ; Male ; Middle Aged ; POEMS Syndrome ; diagnosis ; Syphilis ; diagnosis

Objective To evaluate the clinical application of laparoscopic local resection for gastric tumors.Methods Twenty-three patients with gastric tumors who were performed laparoscopic gastric local resection were retrospectively analyzed with the size of tumor,location of tumor,operative time,blood loss during the operation,time for passage of flatus,post-operative hospital stay,operative complications,post-operative pathological findings and result of follow-up.Results Twenty-three patients were successfully performed laparoscopic local resection,including 8 laparoscopic wedge resection(LWR)and 15 intragastric mucosal resections(IGMR),with no conversion to open surgery.The mean size of gastric tumor was(2.8?1.3)cm,the mean operative time was(82.2?35.5)min,the mean blood loss was(26.5?15.3)mL,the length of incision was(3.1?1.1)cm,the time for passage of flatus was(2.1?0.9)d,and the mean post-operative hospital stay was(7.8?2.0)d.Two patients(8.7%)were found with postoperative gastric mucosal blee-ding and were recovered well through non-operative treatment.The median time of follow-up was 12 months(2-45 months),and no recurrent tumor was observed.Conclusion Laparoscopic local resection is a feasible,safe,effective and less invasive procedure for gastric tumors.

OBJECTIVETo establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA).

METHODSLCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis.

RESULTS8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment.

CONCLUSIONSuccessfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.

Adsorption ; Chromatography, Affinity ; methods ; Immunoelectrophoresis ; Lens Plant ; Plant Lectins ; chemistry ; Reproducibility of Results ; alpha-Fetoproteins ; chemistry ; isolation & purification