Objective To study the treatment effect of endoscopic sinus surgery in the elderly patients with chronic sinusitis and nasal polyps.Methods The results of endoscopic sinus surgery in 60 elder patients were analysed retrospectively.Results The total response rate was 93.4%(56/60),curative rate was 76.7%(46/60),improvement rate was 16.7%(10/60) with over one year followed-up.Conclusion The endoscopic sinus surgery is also applicable to patients with chronic sinusitis and nasal polyps.

Objective To express and purify EGF-TCS fusion protein and observe the targeted and selective killing effect on cancer cells of the fusion protein.Methods The recombinant expression plasmid PQE30/EGF-TCS was transformed to E.coli.M15 and the fusion protein(EGF-TCS) was expressed.Ni-NTA Agrose affinity chromatography was used to purify the protein,flow cytometry to detect EGFR expression rate in cancer cells(BEL-7402,MCF-7,BGC-823) and normal liver LO2 cells,and the killing test to verify selective killing ability of EGF-TCS;The cell apoptosis detection by flow cytometry and microscopic observation were used to confirm the selective killing ability of EGF-TCS.Results Recombinant expression plasmid PQE30/EGF-TCS was expressed in E.coli.M15 stably and effectively.The purity of EGF-TCS was over 95% by chromatography.EGFR expression rate was highest in hepatoma cells BEL-7402(72.33%) and lowest in normal liver LO2 cells(5.51%).The killing ability of recombinant protein was more effective to cancer cells(IC50 of BEL-7402,MCF-7 and BGC-823 was 11.4,22.47 and 12.53 ?g/ml respectively) and was weak to normal cells(IC50 53.19 ?g/ml).Conclusion The recombinant protein EGF-TCS that induces apoptosis of cancer cells was successfully constructed by gene engineering technology.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

In order to investigate the impact of fulvic acid (FA) on the hydroxylysyl glycosylation in collagen bio-synthesis, 40 NMRI mice were divided into two groups (n = 20 in each group, consisting 10 females and 10 males). The animal was maintained for two generations by different diets: control group with normal water and food and study group with water containing 30 mg/L FA and normal food. The second generation of the animal was slaughtered, and the biochemical parameters of collagen content and the degree of collagen hydroxylysyl glycosylation in skin, rib and tibia were detected by biochemical methods. The mean value of collagen in the study group was increased slightly, and no significant difference between study group and control group was found (P > 0.05), but the content of glucose-glactose-hydroxylysine (GGH) was significantly decreased in the study group in comparison with the control group (P<0.01). It was suggested that through the decrease of GGH 30 mg/L FA could inhibit the activity of galactosyl-hydroxylysylglucosyl-transferase and further disturb the post-translational modification of collagen intracellularly.
Benzopyrans/*pharmacology ; Bone Development ; Bone and Bones/chemistry ; Bone and Bones/*metabolism ; Collagen/*biosynthesis ; Glycosylation ; Hydroxylysine/*metabolism ; Mice, Inbred Strains ; Osteoarthritis/etiology ; Selenium/deficiency

Objective: To explore the application value on combined examination of blood levels of growth differentiation factor-15 (GDF-15) and NT-pro B-type natriuretic peptide (NT-proBNP) in patients after successful cardiopulmonary resuscitation (CPR) for their recent prognosis.
Methods: A total of 102 patients with sudden cardiac arrest and successful CPR in our hospital were enrolled. Blood levels of GDF-15 were examined at immediately, 12 h and 24-48 h after CPR respectively. According to GDF-15 levels, the patients were divided into 3 groups: Group A, the patients with GDF-15<1200 ng/L at all-time points,n=31; Group B, GDF-15 level consistently increasing and GDF-15>1200 ng/L at all-time points,n=35; Group C, GDF-15 level consistently increasing at 12 h and 24-48 h after CPR, while it was lower at 24-48 h than 12 h after CPR,n=36. Blood levels of NT-proBNP and left ventricular ejection fraction (LVEF) were also examined. The patients were followed-up for 6 months for post-CPR death.
Results: Blood levels of GDF-15 and NT-proBNP were related, NT-proBNP level was changing with GDF-15 varying. GDF-15 and NT-proBNP level was negatively related to LVEF (r=-0.530,P<0.001), the patients with GDF-15>1800 ng/L and NT-proBNP>400 pg/ml had the higher mortality than those had the lower levels of GDF-15 and NT-proBNP,P<0.05. Survival analysis presented that 6 months survival rate in Group B was lower than Group A and Group C,P<0.05; survival rate was similar between Group A and Group C,P>0.05.
Conclusion: Combined examination for blood levels of GDF-15 and NT-proBNP may better predict the recent prognosis in patients who received CPR.

Objective To express human-mouse chimeric antibody against anthrax protective anti-gen and to analyze its biological activities. Methods A new mammalian bipromoter expression vector was constructed with dihydrofolate reduetase(DHFR) gene as the selection and complication marker. First, the light and heavy chain variable region gene of the monoclonal antibody 5E1 were cloned by RT-PCR, at the same time the human IgG1 heavy chain constant region gene and kappa type constant region gene were cloned. Next, the human-mouse chimeric antibody genes were synthesized by fusion PCR. Then, the hu-man-mouse chimeric antibody gene were inserted into MCS of pSecTag and B1 to construct pSecTag-5E1L and B1-5E1H, respectively. Finally, heavy chain expression cassette excised from the B1-5E1H with Bgl Ⅱ/BamH Ⅰ was further cloned into the Bgl Ⅱ site of the pSecTag-5E1L to construct pSecTag-5E1. Plasmid pSecTag-5E1 was transfected into CHO(dhfr) engineering cells and high production cell clones that were screened by enhancing MTX concentration. After collecting medium and purifying chimeric antibody with af-finity chromatogram, purified chimeric antibody was analyzed by SDS-PAGE, Western blot. Results A sta-ble and high production cell line was acquired at MTX concentration 5×10~(-8) mol/L. Conclusion The hu-man-mouse chimeric antibodies were successfully expressed in CHO cells.

In order to investigate the impact of fulvic acid (FA) on the hydroxylysyl glycosylation in collagen bio-synthesis, 40 NMRI mice were divided into two groups (n = 20 in each group, consisting 10 females and 10 males). The animal was maintained for two generations by different diets: control group with normal water and food and study group with water containing 30 mg/L FA and normal food. The second generation of the animal was slaughtered, and the biochemical parameters of collagen content and the degree of collagen hydroxylysyl glycosylation in skin, rib and tibia were detected by biochemical methods. The mean value of collagen in the study group was increased slightly, and no significant difference between study group and control group was found (P > 0.05), but the content of glucose-glactose-hydroxylysine (GGH) was significantly decreased in the study group in comparison with the control group (P<0.01). It was suggested that through the decrease of GGH 30 mg/L FA could inhibit the activity of galactosyl-hydroxylysylglucosyl-transferase and further disturb the post-translational modification of collagen intracellularly.
Animals ; Benzopyrans ; pharmacology ; Bone Development ; Bone and Bones ; chemistry ; metabolism ; Collagen ; biosynthesis ; Female ; Glycosylation ; Hydroxylysine ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Osteoarthritis ; etiology ; Selenium ; deficiency

Objective:To systematically evaluate the bias risk and applicability of short-term mortality risk prediction models within 30 days after stroke, providing a basis for selecting or developing standardized risk prediction models.Methods:Research on short-term mortality risk prediction models within 30 days after stroke was electronically retrieved from China National Knowledge Infrastructure, WanFang Data, VIP, and China Biomedical Database, PubMed, Web of Science, Embase, Cochrane Library and CINAHL. The search period was from database establishment to December 5, 2023. Two researchers independently conducted literature screening and quality evaluation.Results:Twelve studies were included, and a total of 31 models were internally validated, with 7 models undergoing external validation based on internal validation. 26 models reported discriminative power, and 18 models reported calibration methods. The most frequent predictors of modeling were age, hypertension, atrial fibrillation, diabetes and admission Glasgow Coma Scale score. Due to methodological problems such as insufficient sample size, improper handling of missing variables, and inadequate reporting of modeling information, all included studies were rated as high risk of bias.Conclusions:The research on short-term mortality risk prediction models for stroke patients is still in the development stage. Although it has good applicability, the risk of bias is relatively high. Future research should be designed and reported based on prediction model risk of bias assessment tool (PROBAST) and transparent reporting of a multivariable prediction model for individual prognosis or diagnosis (TRIPOD) to avoid common problems summarized in this study and reduce the risk of bias.

We converted the TGC codon (307-309 bp) of Aspergillus flavus urate oxidase (UOX) gene to a GCC codon by using fusion PCR techniques to produce a C103A mutant. This gene was cloned into expression vector pET-42a (+) and then transformed into Escherichia coli BL21 (DE3). The mutant protein (UOX-Ala103) was expressed in soluble form at high levels after induction with IPTG The expressed rUOX-Ala103 accounted for about 45% of total bacterial proteins, rUOX-Ala103 of up to 98% purity was obtained after purified using hydrophobic interaction and anion exchange. Western blotting showed that the anti-UOX antibody specifically recognized rUOX-Ala103. The mutant protein showed a 60% increased in vitro biological activities compared with native protein, and performed a good activity of degrading the uric acid in vivo.
Aspergillus flavus ; enzymology ; Cloning, Molecular ; Codon ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Mutation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Urate Oxidase ; biosynthesis ; genetics ; isolation & purification ; Uric Acid ; metabolism

Objective To investigate the impact of motilin(MTL), neurotensin(NT)and nitric oxide synthase(NOS)on Oddi sphincter(SO)motion after cholecystectomy. Methods Oddi sphincter manometry(SOM)was performed on both Guinea pig model group(cholecystectomy)and control group (laparotomy)12 weeks after operation. Sphincter of Oddi dysfunction(SOD)group was determined by receiver operating characteristic(ROC)curve analysis and area under curve(AUC). Protein expression of MTL, NT and NOS in SO was also detected through integral optical density method. Meanwhile,the contents of MTL and NT in patients′ plasma of both SOD group(SO pressure> 40 mmHg)and control group were compared. Results AUC of 0.75 and SO pressure of more than 29.8 mmHg was determined as the standard of SOD group.MTL and NT contents(193.16±29.2 pg/mL and 104.57±19.52 pg/mL,respectively)of the model group(n=10)in plasma were significant higher than those of control group(n=11)(154.24 ± 27.69 pg/mL and 79.65±11.24 pg/mL,respectively),and same trend of MTL and NT protein expression in SO was detected(3 556.71±455.80 and 6 321.74±203.54 of the model group;3 075.92±350.06 and 5 843.57±344.00 of the control group).While NOS protein expression in model group was lower than that of the control group(2 954.21± 173.54 VS 3 314.91± 246.67, P<0.05). In clinical research, the plasma contents of MTL(350.98 ± 24.31 pg/mL VS 319.56 ± 23.54 pg/mL)and NT(102.39 ± 19.56 pg/mL VS 80.45±12.35 pg/mL)in SOD group(n=15)were higher than those of the control group(n=15)(P<0.05). Conclusion MTL and NT contents in plasma and protein expression of MTL, NT and NOS in SO may be related to SOD. MTL and NT examinations may assist diagnosing SOD after cholecystectomy.