Therapeutic effects of ranibizumab combined with trabeculectomy and panretinal photocoagulation for neovascular glaucoma / 眼科新进展

Objective To evaluate the clinical efficacy of intravitreal ranibizumab injection and panretinal photocoagulation for neovascular glaucoma.Methods Totally 8 patients (40 eyes) with neovascular glaucoma in the First Affiliated Hospital of Zhengzhou university from January 2016 to October 2016 were collected in this study,and the patients were randomly divided into two groups according to the different treatments:trabeculectomy group (18 eyes),in which the patients were treated with intravitreal injection of ranibizumab + trabeculectomy + panretinal photocoagulation,and cyclophotocoagulation group (22 eyes),subjects undergoing cyclophotocoagulation + panretinal photocoagulation.All patients were followed up for 6 months.And there was no significant difference in the general data of the two groups (all P ＞ 0.05).Finally,variables including iris neovascularization,the best corrected visual acuity (BCVA),the preoperative and postoperative 1 month,3 months and 6 months intraocular pressure,the operation successful rate and complications in both groups were evaluated and compared.Results In the trabeculectomy group,1 week after treatment,neovascularization in all patients was disappeared,and 6 months of follow-up,a small amount of neovascular recurrence occurred;in the cyclophotocoagulation group,2 weeks after treatment,neovascularization disappeared in 18 eyes,decreased in 3 eyes,and did not change in 1 eye;additionally,6 months of follow-up,some eyes presented neovascularization recurrence.The postoperative BCVA in the trabeculectomy group was significantly higher than that before operation,and the difference was statistically significant (P =0.026),But there was no significant difference between preoperative and postoperative BCVA in the cyclophotocoagulation group (P =0.438).Preoperative BCVA in the trabecuiectomy group was significantly higher than that in the cyclophotocoagulation group,and the difference was statistically significant (P =O.039).The intraocular pressures of the both groups were decreased at 1 month,3 months and 6 months after operation,and the differences were statistically significant (all P ＜ 0.05).There was significant difference in the success rate of operation between the two groups (P =0.047).The efficacy of the trabeculectomy group was better than that of the cyclophotocoagulation group.The incidence of complications in the trabeculectomy group was lower than that in the cyclophotocoagulation group,and the difference was statistically significant (P =O.024).Conclusion It is safe and effective for intravitreal ranibizumab injection + trabeculectomy + panretinal photocoagulation in the treatment of iris neovascularization,of which the clinical efficacy is better than cyclophotocoagulation + pamretinal photocoagulation.

Expression of pectin lyase 1 from Aspergillus oryzae in Escherichia coli / 生物工程学报

Pectin lyases from Aspergillus oryzae and Aspergillus niger are usually used for the production of traditional fermented foods, but these fungi produce less pectinases under natural conditions. The cDNA coding mature Pell (without signal peptide) was amplified from Aspergillus oryzae by RT-PCR. Pell cDNA was cloned into pET-28a ( + ) expression vector, then was transformed into E. coli Turner (DE3) plac I cells to express Pell with 6-His tag. For improving the efficiency of Pell expression in E. coli, the conditions of expressing the Pell in E. coli were optimized. E. coli Turner (DE3) plac I cells with pET-28a ( + )-pell was first cultivated at 37 degrees C, 220 r/min until OD600 reached about 0.8. Then, cultivation broth was added with 0.05-0.1 mmol/L IPTG and continuously incubated at 15 degrees C, at 170 r/min for 60 h for expressing of Pell. The recombinant expressed Pell activity could reach 400 u/mL medium, which is 4000-fold of Pell produced naturally by A. oryzae and superior than known recombinant amount of pectin lyases expressed in different fungi expression systems.

Expression of endopolygalacturonase A of Aspergillus oryzae in Escherichia coli / 生物工程学报

Pectinases are mainly used in the food industry to clarify fruit juices and wine, improve oil extraction, remove the peel from the citrus fruit, increase the firmness of some fruits and degum fibres. The filamentous fungus Aspergillus oryzae, used for the production of traditional fermented foods, only could produce less pectinases under general conditions. So far only a few of PGs expressed in yeast or E. coli were reported but they did not show higher activity. The cDNA of mature PGA (without signal peptide) was synthesized with specific primers from total RNA of Aspergillus oryzae by RT-PCR. PGA cDNA was ligated into pET-28a( + ) expression vector, creating plasmid pET-28a( + )-pgA. The plasmid pET-28a( + )-pgA was transformed into E. coli Turner (DE3) plac I cells to express PGA heterogeneously. For improving the efficiency of PGA expression in E. coli, the conditions for expression of the PGA in E. coli were optimized. E. coli Turner (DE3) plac I cells with pET-28a( + )-pgA was first cultivated at 37 degrees, 220r/min until OD600nm reached about 0.8. Then, cultivation broth was added with 0.5 mmol/L IPTG and incubated at 15 degrees C, 170r/min for other 24 h for induced-expression of PGA. Our data showed that the activity of recombinant expressed PGA could reach to 70u/mL medium, which is 87.5-fold of the activity of PGA produced in culture of A. oryzae and superior than known recombinant expression amount of PGA reported by other researchers.

Cloning and expression of xyn III from genomic DNA of Trichoderma reesei QM9414 by overlap-PCR / 生物工程学报

After the cell enters into its programmed cell death, xylanases from grass plants gradually matured through its N-terminal and C-terminal sequence been cut by acid proteases several times. They could not be expressed by conventional protein expression system. Search the GenBank database, xynIII from a mutant of T. reesei QM9414(ATCC26921)was found. It is similar to grass plants' xylanase in their families and structures. It couldn't express in T. reesei QM9414, but its gene exist in genomic DNA as one copy. Through overlap-PCR method, 4 exons of xynIII were cloned, sequenced, spliced, and the whole cDNA of mature xynIII was acquired. The cDNA was inserted into pETBlue-2 vector and transformed into E. coli DE3 pLacI cell. Xyn III could be expressed in the transformed cell under the conditions of 37 degrees C, 1 mmol/L IPTG induced for 3h. Low temperature (15 degrees C), long time(64h) induction(0.2 mmol/L IPTG) could enhance xynIII activity.

Study on the immuno-effects of recombinant hepatitis B vaccination in adults / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate the immuno-effects of hepatitis B (HB) vaccination in adults.</p><p><b>METHODS</b>Five groups were sampled by means of cluster sampling, and serum HBsAg, anti-HBs and anti-HBc were tested in every group at people aged from 18 to 50. Recombinant HB vaccine was injected to the ones that HBsAg, anti-HBs and anti-HBc were all negative. Concentration of anti-HBs in serum was tested after one year and three years of vaccination. Immuno-effects of recombinant HB vaccination in adults at different ages and between sexes, were then calculated.</p><p><b>RESULTS</b>Good immuno-effects of recombinant HB vaccination in adults were noticed. After one year and three years of vaccination with 5 micro g recombinant HB vaccine, the anti-HBs positive rates were 82.76%, 70.77% while the serum concentrations of anti-HBs were 55.91 mIU/ml and 35.41 mIU/ml respectively. When 10 micro g was used, the concentrations were 83.74%, 72.22%, 56.89 mIU/ml and 30.29 mIU/ml respectively. The effects did not show significant differences between different doses on 10 micro g and of 5 micro g. Concentration of anti-HBs reduced when time went by. The factors such as age and sex influenced the effects of immunity on recombinant HB vaccination.</p><p><b>CONCLUSION</b>Good immunity could be obtained when recombinant hepatitis B was vaccinated in vulnerable population aged 18 to 50.</p>