1.Prokaryotic expression of Staphylococcus aureus Clumping factor B and evaluation of the antiserum-mediated opsonic activity.
Lili ZHANG ; Shanshan CAO ; Yifeng ZHANG ; Chunling XUAN ; Youqiang WU ; Songhua HU
Chinese Journal of Biotechnology 2015;31(11):1623-1631
Staphylococcus aureus is a major cause of hospital-acquired infection. Because the bacteria are very easy to become resistant to antibiotics, vaccination is a main method against S. aureus infection. Clumping factor B (ClfB) is an adhesion molecule essential for S. aureus to colonize in the host mucosa and is regarded as an important target antigen. In this study, we successfully used Escherichia coli to express a segment encoding the N1-N3 regions of ClfB protein (Truncated-ClfB) cloned from S. aureus. The protein was purified by affinity and ion exchange chromatographies and gel filtration. Rabbits were immunized three times with purified Truncated-ClfB. After that, blood was collected to prepare serum which were then used for measurement of antibody level. Phagocytosis of S. aureus opsonized by the serum was determined by a flow cytometry. Results show that the serum IgG titer reached 1:640 000. Phagocytosed S. aureus by polymorphonuclear leukocytes were significantly more when the bacteria were opsonized by the serum from Truncated-ClfB immunized rabbits than those from no immunized group (P < 0.01). Therefore, the results indicated that Truncated-ClfB could be a promising vaccine candidate against S. aureus infection.
Adhesins, Bacterial
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immunology
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Animals
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Antibodies, Bacterial
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blood
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Escherichia coli
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Flow Cytometry
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Immune Sera
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Immunoglobulin G
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blood
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Opsonin Proteins
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immunology
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Phagocytosis
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Rabbits
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Staphylococcal Infections
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immunology
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Staphylococcus aureus
2.Direct trocar insertion using optical trocar in the establishment of the observational passage during trans-peritoneal laparoscopic operation-our initial experience
Jieying WU ; Jinming DI ; Yun LUO ; Shrestha RUJAN ; Youqiang FANG ; Cheng HU
Chinese Journal of Urology 2016;37(8):586-590
Objective To evaluate the clinical significance of direct trocar insertion using optical trocar in the establishment of the primary port during trans-peritoneal laparoscopic surgical procedures.Methods A prospective study was conducted by collecting the data of 120 patients who should be performed abdominal laparoscopic surgery from April 2015 to December 2015.The 120 patients were randomly divided into a research group and a control group.The research group consisted of 34 male patients and 26 female patients,mean age was (52.0 ± 11.9) years and mean BMI was (24.9 ± 2.9) kg/m2.In research group,patients were positioned laterally with the flank padded and elevated.A predetermined position was drawn prior to surgery between the umbilicus and lateral rectus abdominis,for the creation of the primary laparoscopic trocar port.The predetermined point was incised,and then the method of direct trocar insertion using the optical access trocar was used for establishment of the primary port.After this maneuver was completed the surgery continued as indicated.The control group consisted of 36 male patients and 24 female patients,whose mean age was (52.9 ± 11.4) years and mean BMI was (25.2 ± 2.4) kg/m2.This group underwent the traditional method of port construction by incision into the abdomen.The time of constructing the passage,leakage rate,bleeding rate,and injury rate of abdominal organs were compared.Results In research group,the time of building primary port was clearly shorter than that in control group (2.7min vs.15.9min,P < 0.05),the leakage rate was also obviously reduced compared to that in control group (0 vs.30%,P < 0.05).Neither groups observed any significant bleeding nor visceral organ damage throughout the study.Conclusion Direct trocar insertion using optical trocar to establish observation port is a highly efficient and safe method in trans-peritoneal laparoscopic operation,which should be research thoroughly in clinical practice.
3.Helix hydro-jet compared with the holmium laser lithotripsy in the treatment of intrahepatic bile duct stones
Jianshen LIANG ; Youqiang YE ; Cong TANG ; Yiwu WU ; Zhiping TANG ; Bingzong HOU
Chinese Journal of General Surgery 2014;29(7):514-516
Objective To evaluate the curative effect,safety,advantages and disadvantages of helix hydro-jet and holmium laser lithotripsy in the treatment of intrahepatic bile duct stones.Methods 158 patients of intrahepatic bile duct stones were divided into helix hydro-jet group (n =82) and helix hydro-jet group (n =76).Time of lithotripsy,frequencies of lithotripsy,residual stone,intraoperative hemorrhage,postoperative hemorrhage,stone recurrence rate,postoperative fever,and therapeutic efficacy were compared between the two groups.Results There was no significant difference between the two groups on frequency of lithotripsy,residual stone rate and stone recurrence rate (P > 0.05) ; whereas intraoperative washing normal saline (2 300 ± 275) ml vs (2 850 ± 331) ml,(P =0.002),the time of lithotripsy (54.2 ± 15.3) min vs (38.1 ± 12.7) min,P =0.041,intraoperative hemorrhage (19 cases vs 29 cases,P =0.041),Intraoperative analgesia (2 cases vs 9 cases,P =0.032),postoperative hemorrhage (11 cases vs 21 cases,P =0.037),and postoperative fever (30 cases vs 62 cases,P =0.027) had significant differences(P < 0.05).Conclusions Both helix hydro-jet and the holmium laser lithotripsy is safe and effective for the treatment of intrahepatic bile duct stones.Holmium laser is more efficient than Helix Hydro-jet,while helix hydro-jet is safer than holmium laser in lithotripsy.
4.Construction of Saccharomyces cerevisiae mutant with knockout of SNF4 gene.
Xiaohua LIN ; Chongrong KE ; Bisha WU ; Yongbiao ZHENG ; Li LI ; Youqiang CHEN ; Jianzhong HUANG
Chinese Journal of Biotechnology 2011;27(4):572-578
Construction and ethanol production effects of SNF4 gene knockout in Saccharomyces cerevisiae were described in this paper. For knockout of SNF4 gene in S. cerevisiae YS2, a PCR-amplified disruption cassette was used, encoding the short flanking homologous regions to the SNF4 gene and Kan(r) as selectable marker. The SNF4 gene disruption cassette was transformed into S. cerevisiae YS2 through LiAc/SS Carrier DNA/PEG. The positive transformants were grown on G418 plates and verified by PCR. The Kan(r) marker was rescued by transforming plasmid pSH65 into positive transformants and inducing expression of Cre recombinase in galactose-containing medium. Lastly, the YS2-deltaSNF4 strain, in which SNF4 allele gene were completely knocked out, was obtained by repeating the same procedure. The result of anaerobic fermentation showed that ethanol production of the SNF4 gene knockout strain had increased by 7.57 percent as compared with the original strain YS2. The experiment indicated ethanol production could be improved significantly with the approach ofSNF4 gene knockout by Cre-LoxP system.
AMP-Activated Protein Kinases
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genetics
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Ethanol
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metabolism
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Fermentation
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Gene Knockout Techniques
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methods
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Mutation
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Saccharomyces cerevisiae
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genetics
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Saccharomyces cerevisiae Proteins
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genetics
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Transcription Factors
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genetics
5.Effects of Vestibular Function Rehabilitation Combined with Proprioceptive Sense Training on Senile Posterior Circulation Ischemic Vertigo
Ruwei XU ; Youqiang FAN ; Di WU ; Peng JIANG ; Lin XU ; Liang XU ; Ming MA ; Yana JIANG
Chinese Journal of Rehabilitation Theory and Practice 2018;24(11):1333-1337
Objective To observe the effects of vestibular function training and proprioception training on vertigo and balance function after postoperative circulatory ischemia under the visual participation. Methods From June, 2016 to May, 2018, 72 patients with posterior circulation ischemic vertigo aged more than 60 were randomly divided into proprioception group (n = 24), vestibular group (n = 24) and comprehensive group (n = 24). All the patients received routine medicine and rehabilitation training. Moreover, the proprioception group accepted proprioception training, the vestibular group accepted vestibular function training, and the comprehensive group accepted both proprioception training and vestibular function training. They were assessed with Dizziness Assessment Rating Scale (DARS), Dizziness Handicap Inventory (DHI) and Berg Balance Scale (BBS) before and after four weeks of treatment, while the mean velocity (Vm) of blood flow in the basilar artery was measured with transcranial Doppler, and the stability index (ST) was measured with Tetrax. Results The scores of DARS, DHI and BBS, and Vm and ST improved in all the groups after treatment (t > 57.825, P < 0.001). However, the scores of DHI and BBS, and ST improved more in the comprehensive group than in the proprioception group and the vestibular training group (P < 0.05), Vm and score of DARS improved more in the vestibular group and the comprehensive group than in the proprioception group (P < 0.05). Conclusion With the participation of the vision, vestibular function training combined with proprioception training can further alleviate vertigo caused by postoperative circulatory ischemia, and improve balance and quality of life.