The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.

Objective:To optimize the formula of film-forming materials for compound rhizoma corydalis plastics. Methods:Poly-vinyl alcohol 124 (PVA124) and bletilla striata gum were selected as the film-forming materials. The amounts of PVA 124, bletilla striata gum, glycerol and absolute ethanol were used as the influencing factors in an orthogonal test with the composite score of film-forming time and appearance as the indices to screen out the best composition of film-forming materials. Results:The best composition of film-forming materials was as follows:8 g PVA 124, 1 g bletilla striata gum, 2 ml glycerol and 15 ml ethanol. Conclusion:The op-timized film-forming materials can be used for the preparation of compound rhizoma corydalis plastics.

The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an α-keto group and a β-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts.
Bacterial Proteins ; genetics ; metabolism ; Catalysis ; Cyclohexanones ; metabolism ; Escherichia coli ; enzymology ; Polyketide Synthases ; genetics ; metabolism ; Protein Structure, Tertiary ; Substrate Specificity ; omega-Chloroacetophenone ; metabolism

Purpose: To report a case of contact lens-related infectious keratitis caused by three different bacterial species.Case summary: A 40-year-old man presented with pain and redness in his left eye. His best-corrected visual acuity (BCVA) in the affected eye was 20/25. Slit-lamp examination revealed a 3 × 3 mm corneal epithelial defect with infiltration located 1 mm inferior to the pupil. Following admission, a microbial culture test was performed, and empirical antibiotic therapy was initiated. On the fourth day, Pseudomonas aeruginosa was isolated from the corneal sample and the contact lens, while Serratia marcescens and Stenotrophomonas maltophilia were isolated from the contact lens case. Based on the results of the antibiotic susceptibility tests, 0.5% moxifloxacin, fortified amikacin, and ceftazidime were administered topically and intravenously. The corneal epithelial defect reduced to 1 × 1 mm by the eleventh day of admission. After two months, BCVA improved to 20/20 with no remaining corneal epithelial defect, although an inactive corneal opacity persisted at the previous ulcer site. Conclusions Contact lens wear can be associated with polymicrobial keratitis involving three distinct Gram-negative bacteria, which may present greater treatment challenges compared to monomicrobial keratitis. Microbial culture testing of the contact lens, its case, and corneal scrapings is essential for identifying the causative organisms and selecting appropriate antibiotic therapy.

Purpose: To report a case of contact lens-related infectious keratitis caused by three different bacterial species.Case summary: A 40-year-old man presented with pain and redness in his left eye. His best-corrected visual acuity (BCVA) in the affected eye was 20/25. Slit-lamp examination revealed a 3 × 3 mm corneal epithelial defect with infiltration located 1 mm inferior to the pupil. Following admission, a microbial culture test was performed, and empirical antibiotic therapy was initiated. On the fourth day, Pseudomonas aeruginosa was isolated from the corneal sample and the contact lens, while Serratia marcescens and Stenotrophomonas maltophilia were isolated from the contact lens case. Based on the results of the antibiotic susceptibility tests, 0.5% moxifloxacin, fortified amikacin, and ceftazidime were administered topically and intravenously. The corneal epithelial defect reduced to 1 × 1 mm by the eleventh day of admission. After two months, BCVA improved to 20/20 with no remaining corneal epithelial defect, although an inactive corneal opacity persisted at the previous ulcer site. Conclusions Contact lens wear can be associated with polymicrobial keratitis involving three distinct Gram-negative bacteria, which may present greater treatment challenges compared to monomicrobial keratitis. Microbial culture testing of the contact lens, its case, and corneal scrapings is essential for identifying the causative organisms and selecting appropriate antibiotic therapy.

Purpose: To report a case of contact lens-related infectious keratitis caused by three different bacterial species.Case summary: A 40-year-old man presented with pain and redness in his left eye. His best-corrected visual acuity (BCVA) in the affected eye was 20/25. Slit-lamp examination revealed a 3 × 3 mm corneal epithelial defect with infiltration located 1 mm inferior to the pupil. Following admission, a microbial culture test was performed, and empirical antibiotic therapy was initiated. On the fourth day, Pseudomonas aeruginosa was isolated from the corneal sample and the contact lens, while Serratia marcescens and Stenotrophomonas maltophilia were isolated from the contact lens case. Based on the results of the antibiotic susceptibility tests, 0.5% moxifloxacin, fortified amikacin, and ceftazidime were administered topically and intravenously. The corneal epithelial defect reduced to 1 × 1 mm by the eleventh day of admission. After two months, BCVA improved to 20/20 with no remaining corneal epithelial defect, although an inactive corneal opacity persisted at the previous ulcer site. Conclusions Contact lens wear can be associated with polymicrobial keratitis involving three distinct Gram-negative bacteria, which may present greater treatment challenges compared to monomicrobial keratitis. Microbial culture testing of the contact lens, its case, and corneal scrapings is essential for identifying the causative organisms and selecting appropriate antibiotic therapy.

Purpose: To report a case of contact lens-related infectious keratitis caused by three different bacterial species.Case summary: A 40-year-old man presented with pain and redness in his left eye. His best-corrected visual acuity (BCVA) in the affected eye was 20/25. Slit-lamp examination revealed a 3 × 3 mm corneal epithelial defect with infiltration located 1 mm inferior to the pupil. Following admission, a microbial culture test was performed, and empirical antibiotic therapy was initiated. On the fourth day, Pseudomonas aeruginosa was isolated from the corneal sample and the contact lens, while Serratia marcescens and Stenotrophomonas maltophilia were isolated from the contact lens case. Based on the results of the antibiotic susceptibility tests, 0.5% moxifloxacin, fortified amikacin, and ceftazidime were administered topically and intravenously. The corneal epithelial defect reduced to 1 × 1 mm by the eleventh day of admission. After two months, BCVA improved to 20/20 with no remaining corneal epithelial defect, although an inactive corneal opacity persisted at the previous ulcer site. Conclusions Contact lens wear can be associated with polymicrobial keratitis involving three distinct Gram-negative bacteria, which may present greater treatment challenges compared to monomicrobial keratitis. Microbial culture testing of the contact lens, its case, and corneal scrapings is essential for identifying the causative organisms and selecting appropriate antibiotic therapy.

PURPOSE: To investigate the change of refractive error between the full-correction and under-correction treatment groups of myopic anisometropic patients. METHODS: This study included 36 patients who had no amblyopia with myopic anisometropia > 3.00 diopters (D) and less than 6.00 D using the cycloplegic refraction test. The patients were divided into two groups involving the full-correction of both eyes (group 1) or full-correction on the less myopic eye and under-correction with −0.50 D of the more myopic eye (group 2). We monitored refractive changes every 6 months for 24 months. RESULTS: At the first visit, the mean refractive error of the less myopic eye was −0.68 ± 0.54 D and that of the more myopic eye was −4.22 ± 0.77 D in group 1. The mean refractive error of the less myopic eye was −0.75 ± 0.58 D and that of the more myopic eye was −4.36 ± 0.73 D in group 2. There was no significant difference between the groups (p = 0.713 and p = 0.585, respectively). At 24 months, the mean refractive errors of group 1 were −1.27 ± 0.54 D and −4.88 ± 0.81 D, respectively, and that of group 2 were 1.38 ± 0.54 D and −5.59 ± 1.01 D, respectively. The mean refractive error of the less myopic eyes showed no significant difference between both groups (p = 0.555), but that of the more myopic eyes was significantly different (p = 0.027). Between both groups, the degree of anisometropia at 24 months was 3.61 ± 0.60 in group 1 and 4.20 ± 0.86 in group 2. Group 2 showed a significant difference and more severe anisometropic changes (p = 0.022). CONCLUSIONS: Full correction of myopic anisometropia without amblyopia is a better method for reducing the progression of anisometropia.
Amblyopia ; Anisometropia ; Humans ; Methods ; Myopia ; Refractive Errors

OBJECTIVETo introduce practical diagnostic criterion of blood stasis syndrome (BSS), and to evaluate its reliability and validity.

METHODSBy referring to three diagnostic criteria of BSS [practical diagnostic criterion of BSS (criterion A), diagnostic criterion of BSS in 1986 (criterion B), Consensus of Integrative Medicine on BSS Diagnosis in 2011 (criterion C)], 712 patients from different departments of Xiyuan Hospital were recruited. The reliability of criterion A and its consistency with the other two criteria were assessed using Kappa coefficient. A Bayesian approach was also employed to assess the sensitivity and specificity of criterion A.

RESULTSAccording to the consistency check, criterion A presented good consistency when used by different researchers (the diagnostic accordance rate was 91. 96%, Kappa =0. 82, P <0.001). Meanwhile, there was an acceptable diagnostic consistency among the three diagnostic criteria. Bayesian estimation suggested that criterion A had higher sensitivity but similar specificity, as compared with criterion B or criterion C. Compared with criterion B [the median of sensitivity and specificity were 0. 762 (95% Cl: 0. 731 -0. 790) and 0. 902 (95% Cl: 0. 858 -0. 936) respectively, the median of sensitivity and specificity of criterion A were 0. 911 (95% CI: 0. 888 - 0. 930) and 0. 875 (95% CI: 0. 826 - 0. 915) respectively. Estimating the difference between criterion A and B, the median of sensitivity and specificity were 0. 149 (95% CI: 0. 112 -0.184) and -0. 026 (95% CI:-0. 085 -0. 033) respectively. Compared with criterion C [the median of sensitivity and specificity were 0. 831 (95% Cl: 0. 804 -0. 857) and 0. 892 (95% CI: 0. 848 - 0. 926) respectively], the median of sensitivity and specificity of criterion A were 0. 912 (95% CI: 0. 889 -0. 932) and 0. 880 (95%CI: 0. 833 - 0.919) respectively. Estimating the difference between criterion A and C, the median of sensitivity and specificity were 0. 081 (95% CI: 0.047 - 0.114) and -0.011 (95%CI: -0.070 -0.046) respectively.

CONCLUSIONCompared with criterion B and C, criterion A not only had better reliability, but also could significantly improve the sensitivity without obviously lowering the specificity.

Objective To investigate the efficacy of interferon α(IFNα)and adefovir dipivoxil (ADV)combination therapy in HBeAg positive chronic hepatitis B(CHB)patients and to explore the optimized strategy for individualized treatment.Methods A total of 156 HBeAg positive CHB patients were enrolled in the study from January 2005 to June 2009 in the Third Affiliated Hospital of Hebei Medical University.Fifty-six CHB patients with hepatitis B virus(HBV)DNA≥1 X 107copy/mLand/or liver fibrosis stage≥S3,or previous monotherapy failure(relapse)were treated with initial IFNα and ADV combination therapy.Fifty-two patients who didn't meet any of the above baseline characteristics received initial IFNα monotherapy.The remaining 48 patients treated with IFNα monotherapy for full treatment duration were considered as control.At week 24 of treatment,the treatment regimens were adjusted according to quantitative changes of HBV DNA,HBeAg and HBsAg:16 patients who achieved early response in group of initial IFNα and ADV combination therapy subsequently received IFNα monotherapy,the other patients in group of initial combination therapy together with patients who did not achieved early response in group of initial IFNα monotherapy subsequently received IFNα and ADV combination treatment.The HBV DNA levels,HBeAg and HBsAg titers were detected at the end of 48 weeks of treatment to determine the treatment duration.The treatment efficacy,safety,drug resistance and relapse rates were finally evaluated at week 72.All data were analyzed using chi square test.Results At week 24,the early response rate in group of initial combination therapy was 28.6%,and the HBV DNA negative rate and alanine aminotransferase(ALT)normalization rate were significantly higher than those in groups of initial IFNα monotherapy and control(53.6%vs 32.7%vs 27.1%and 62.5%vs 40.4%vs 37.5%,respectively,P<0.05);in addition,HBeAg loss rate was higher than control group(39.3%vs 18.8%,x2=7.48;P<0.05).At week 48,five of 16 patients who achieved early response developed HBeAg reversion and three cases accompanied with virological breakthrough in group of initial combination therapy after switching to IFNα monotherapy,while the rates of HBV DNA negative,HBeAg seroconversion and HBsAg clearance were 73.2%,41.1%and 12.5%,respectively.The HBV DNA negative rate,HBeAg seroconversion rate and HBsAg clearance rate in 96 patients Who had received different combination treatment regimens were 65.6%,33.3%and 8.3%,respectively.At week 72,the relapse rate in individualized treatment group was comparable to those in control group,while HBsAg clearance rate increased 2.7%in individualized treatment group.Conclusions IFNα and ADV combination treatment could improve early biochemical and virological responses.Individualized treatment strategy based on baseline characteristics and treatment responses may be helpful for optimizing antiviral treatment in CHB patients.