Rigid Gas-Permeable Contact Lens for Visual Rehabilitation in Children Younger Than 12 Years With Penetrative Ocular Trauma.

OBJECTIVES: To observe the clinical outcomes of visual rehabilitation using rigid gas-permeable contact lenses (RGPCLs) after penetrative ocular trauma in children younger than 12 years in China. METHODS: Patients younger than 12 years with penetrative ocular trauma fitted with an RGPCL for visual rehabilitation from 2017 to 2021 were included. In the case cohort, the best-corrected visual acuity (BCVA) with spectacles was measured when the RGPCL was fitted, and the initial BCVA with RGPCL, and the BCVA at the last visit were compared. RESULTS: Fifteen patients, aged 4 to 12 (mean 8.0±2.7) years, who wore an RGPCL for 7 to 53 (mean 20.3±15.7) months, were included. The BCVA was log of minimal angle of resolution 0.4 (0.2-0.7) with spectacles and 0.1 (0.1-0.2) for RGPCL at the initial visit, and 0.0 (0.0-0.1) for BCVA at the last visit, with a statistically significant difference between the three comparisons ( P <0.001). Six of the 15 (40%) children abandoned wearing RGPCL because of discomfort and lens rejection (n=3, 50%), lens loss and inability to replace broken lens because of travel distances and epidemics (n=2, 33%), and cost (n=1, 17%). CONCLUSIONS: Although application is complicated and initial wearing comfort is poor, an RGPCL is still a beneficial, safe tool for postoperative visual rehabilitation in children with open ocular trauma.

Natural Nano-Drug Delivery System in Coptidis Rhizoma Extract with Modified Berberine Hydrochloride Pharmacokinetics.

PURPOSE: This study aimed to evaluate the pharmaceutical and pharmacokinetic effects of the natural nanoparticles (Nnps) isolated from Coptidis Rhizoma extract on berberine hydrochloride (BBR) and systematically explore the related mechanisms. METHODS: Firstly, Nnps were isolated from Coptidis Rhizoma extract and then an Nnps-BBR complex was prepared. After qualitative and quantitative analysis in terms of size, Zeta potential, morphology, and composition of the Nnps and the Nnps-BBR complex, the effects of the Nnps on the crystallization of BBR were characterized. The effects of the Nnps on the solubility and dissolution of BBR were then evaluated. In addition, the effects of the Nnps on BBR in terms of cellular uptake, transmembrane transport, metabolic stability, and pharmacokinetics in mice were studied. RESULTS: The Nnps had an average size of 166.6 ± 1.3 nm and Zeta potential of -12.5 ± 0.2 mV. The Nnps were formed by denaturation of co-existing plant proteins with molecular weight < 30 kDa. The Nnps adsorbed or dispersed BBR, thereby promoting BBR transformation from crystal to amorphous form and improving its solubility and dissolution. The Nnps carried and promoted BBR uptake by human colonic adenocarcinoma (Caco-2) cells via caveolae-mediated endocytosis, reducing P-gp-mediated efflux of BBR in mice gut sacs and Madin-Darby canine kidney cells stably expressing the transporter P-gp (MDCK-MDR1) cells. Moreover, the Nnps improved BBR metabolic stability in mouse intestinal S9, promoting BBR intestinal absorption in mice, as shown by increased peak BBR concentration (Cmax, 1182.3 vs 310.2 ng/mL) and exposure level (AUC0-12 h, 2842.8 vs 1447.0 ng·h/mL) in mouse portal vein. In addition, the Nnps increased BBR exposure level in mouse livers (95,443.2 vs 43,586.2 ng·h/g liver). CONCLUSION: The proteinaceous nanoparticles isolated from Coptidis Rhizoma extract can form a natural nano-drug delivery system with BBR, thereby significantly improving the pharmacokinetics of oral BBR.

[Intervention with Schistosoma japonicum cysteine protease inhibitor for treatment of lipopolysaccharide-induced sepsis in mice].

OBJECTIVE: To observe the effect of Schistosoma japonicum cysteine protease inhibitor (rSjCystatin) for treatment of lipopolysaccharide (LPS)-induced sepsis in mice. METHODS: After a week of adaptive feeding, 54 BALB/c mice were randomly divided into normal control group (group A), sepsis group (group B), and rSjCystatin intervention group (group C). The mice in group A received an intraperitoneal injection of PBS (100 µL), and those in groups B and C were injected with PBS (100 µL) containing LPS (10 mg/kg); the mice in group C were also intraperitoneally injected with 25 µg sjCystatin in 100 µL PBS 30 min after LPS injection. From each group, 10 mice were randomly selected 24 h after PBS or LPS injection for detecting serum levels of TNF-α, IL-6, and IL-10 using ELISA and the levels of ALT, AST, BUN, and Cr using automatic biochemical analyzer; the pathological changes in the liver, lung and kidney were observed with HE staining. The remaining 8 mice in each group were used for observing the changes in the general condition and the 72-h survival. RESULTS: The 72-h survival rates of the mice was 100% in group A, 0 in group B, and 36% in group C, showing a significant difference among the 3 groups (P<0.05). Compared with those in group A, the mice in group B exhibited obvious liver, lung, and renal pathologies with increased levels of ALT, AST, BUN, Cr, IL-6, and TNF-α (P<0.05). Treatment with sjCystatin significantly lessened LPS-induced organ pathologies, lowered the levels of liver and renal functional indexes and the pro-inflammatory cytokines, and increased the serum level of IL-10 in the mice (P<0.05). CONCLUSION: SjCystatin can produce a significant therapeutic effect on sepsis induced by LPS in mice.

[Component analysis of excretory/secretory protein from Trichinella spiralis adult worm].

Objective: To analyze the component of adult worm excretory/secretory protein(AWESP) from Trichinella spiralis using the shotgun method, and find out the active component underlying its regulatory effect on colitis in humans. Methods: The T. spiralis AWESP was prepared, separated by SDS-PAGE, lysed with trypsin, and analyzed by shotgun LC-MS/MS. The protein components were determined with the Masco software and classified using the Gene Ontology(GO) method in cellular components, molecular functions, and biological processes. Results: The AWESPs isolated by SDS-PAGE had a Mr of 15 000-116 000. A total of 280 proteins were revealed by LC-MS/MS, of which 96 were identified by Masco software, 98 were putative, and the remaining 86 were unclear. Preliminary results showed that 4 proteins had regulatory potential for colitis, including cysteine protease inhibitor, serine protease, 53 000 excretory/secretory antigen, and glutathione-S-transferase. GO enrichment analysis showed that the identified proteins had 104 different molecular functions, involved in 363 biological processes. Conclusion: As revealed by the Masco software, T. spiralis AWESP has complex components and 96 have been identified in this study. Four of them are preliminarily shown to be associated with the anti-colitis effect of T. spiralis.

Protective effects of parecoxib on rat primary astrocytes from oxidative stress induced by hydrogen peroxide.

OBJECTIVE: To investigate the protective effects of parecoxib from oxidative stress induced by hydrogen peroxide (H2O2) in rat astrocytes in vitro. METHODS: All experiments included 4 groups: (1) negative control (NC) group, without any treatment; (2) H2O2 treatment group, 100 µmol/L H2O2 treatment for 24 h; (3) and (4) parecoxib pretreatment groups, 80 and 160 µmol/L parecoxib treatment for 24 h, respectively, and then treated with 100 µmol/L H2O2. Several indices were investigated, and the expressions of Bax, Bcl-2, and brain-derived neurotrophic factor (BDNF) were quantified. RESULTS: Compared to the NC group, exposure to H2O2 resulted in significant morphological changes, which could be reversed by pretreatment of parecoxib. In addition, H2O2 treatment led to loss of viability (P=0.026) and increased intracellular reactive oxygen species (ROS) levels (P<0.001), and induced apoptosis (P<0.01) in the primary astrocytes relative to the NC group. However, in the parecoxib pretreatment groups, all the above changes reversed significantly (P<0.05) as compared to the H2O2 treatment group, and were nearly unchanged when compared to the NC group. Mechanical investigation showed that dysregulated Bax, Bcl-2, and BDNF could be implicated in these changes. CONCLUSIONS: Our results indicated that parecoxib provided a protective effect from oxidative stress induced by exposure to H2O2.

[In vitro killing of Trichinella spiralis muscle larvae by exogenous nitric oxide from SNP].

OBJECTIVE: To study the lethal effect of exogenous nitric oxide donor sodium nitroprusside (SNP) on the muscle larvae of Trichinella spiralis in vitro cultivation. METHODS: T. spiralis muscle larvae isolated from the infected BALB/c mice were formulated into a 1,000 larva/ml suspension with RPMI 1640 medium, and 0.1 ml suspension per orifice was cultured with SNP at 37°C in a humidified 5% CO2 atmosphere. The final concentrations of SNP were 0.02, 0.05, 0.10, 0.20, 0.50 and 1.00 mmol/L, respectively, and then the experiments were divided into 5 groups:1.00 mmol/L SNP (control group, Group A), 0.15 mmol/L FeSO4+ 1.00 mmol/L SNP (Group B), 1.00 mmol/L L-cysteine + 1.00 mmol/L SNP (Group C), 0.15 mmol/L FeSO4+ 1.00 mmol/L L-cysteine + 1.00 mmol/L SNP (Group D) and 0.15 mmol/L Hemoglobin + 1.00 mmol/L SNP (Group E). All the groups were incubated with T. spiralis muscle larvae in RPMI 1640 medium. The survivability of the muscle larvae was observed by steromicroscope and the differences of inhibition ratio among these groups were analyzed 4 d after the incubation. Results SNP 0.02 mmol/L was not cytotoxic to the muscle larvae with an inhibition of (5.50 ± 1.80) %. The mortality rates of SNP 0.05, 0.10, 0.20, 0.50, 1.00 mmol/L groups were (20.19±2.71)%, (29.21±2.12)%, (41.81±2.03)%, (47.85±3.79)%, (60.98±5.19)%, respectively, significantly higher than that of the control group[(4.93±0.25) %, all P < 0.051]. There was a positive liner correlation between the mortality of muscle larvae and SNP concentrations in the range of 0.02-1.00 mmol/L. Next, Group A, B, C, D and E led to the mortalities from (60.98±5.19)% to (49.48±1.34)%, (47.29±2.79)%, (26.28±1.37)%, (17.93±3.49)%, respectively, and all the differences between Group A and the other four groups were statistically significant (all P < 0.05). CONCLUSIONS: Exogenous nitric oxide released from SNP can kill the muscle larvae of T. spiralis. However, hemoglobin, L-cysteine, and FeSO4 can reverse the lethal effect on the parasites. The best inhibitor was hemoglobin.

Optimized codon usage enhances the expression and immunogenicity of DNA vaccine encoding Taenia solium oncosphere TSOL18 gene.

Cysticercosis due to larval cysts of Taenia solium, is a serious public health problem affecting humans in numerous regions worldwide. The oncospheral stage-specific TSOL18 antigen is a promising candidate for an anti-cysticercosis vaccine. It has been reported that the immunogenicity of the DNA vaccine may be enhanced through codon optimization of candidate genes. The aim of the present study was to further increase the efficacy of the cysticercosis DNA vaccine; therefore, a codon optimized recombinant expression plasmid pVAX1/TSOL18 was developed in order to enhance expression and immunogenicity of TSOL18. The gene encoding TSOL18 of Taenia solium was optimized, and the resulting opt-TSOL18 gene was amplified and expressed. The results of the present study showed that the codon-optimized TSOL18 gene was successfully expressed in CHO-K1 cells, and immunized mice vaccinated with opt-TSOL18 recombinant expression plasmids demonstrated opt­TSOL18 expression in muscle fibers, as determined by immunohistochemistry. In addition, the codon-optimized TSOL18 gene produced a significantly greater effect compared with that of TSOL18 and active spleen cells were markedly stimulated in vaccinated mice. 3H-thymidine incorporation was significantly greater in the opt-TSOL18 group compared with that of the TSOL18, pVAX and blank control groups (P<0.01). In conclusion, the eukaryotic expression vector containing the codon-optimized TSOL18 gene was successfully constructed and was confirmed to be expressed in vivo and in vitro. The expression and immunogenicity of the codon-optimized TSOL18 gene were markedly greater compared with that of the un-optimized gene. Therefore, these results may provide the basis for an optimized TSOL18 gene vaccine against cysticercosis.

Complexation of Al(III) with reduced glutathione in acidic aqueous solutions.

The complexation of reduced glutathione (GSH) in its free and Al(III)-bound species in acidic aqueous solutions was characterized by means of multi-analytical techniques: pH-potentiometry, multinuclear ((1)H, (13)C and (27)Al) and two-dimensional nuclear Overhauser enhancement NMR spectroscopy ((1)H, (1)H-NOESY), electrospray mass spectroscopy (ESI-MS), and ab initio electronic structure calculations. The following results were found. In the 25 degrees C 0.1M KCl and 37 degrees C 0.15M NaCl ionic medium systems, Al(3+) coordinates with the important biomolecule GSH through carboxylate groups to form various mononuclear 1:1 (AlHL, AlH(2)L and AlH(-1)L), 1:2 (AlL(2)) complexes, and dinuclear (Al(2)H(5)L(2)) species, where H(4)L(+) denotes totally protonated GSH. Besides the monodentate complexes through carboxylate groups, the amino groups and the peptide bond imino and carbonyl groups may also be involved in binding with Al(3+) in the bidentate and tridentate complexes. The present data reinforce that the glycine carboxylate group of GSH has a higher microscopic complex formation constant than gamma-glutamyl carboxylate. Compared with simple amino acids, the tripeptide GSH displays a greater affinity for the Al(3+) ion and thus may interfere with aluminum's biological role more significantly.

Speciation of aluminum(III) complexes with oxidized glutathione in acidic aqueous solutions.

The structural speciation aspects, including the binding sites, species, complexation abilities and effects of the oxidized glutathione (GSSG) with aluminum(III) in aqueous solutions, have been studied by means of many analytical techniques: pH-potentiometry (25 degrees C, 0.1 M KCl and 37 degrees C, 0.15 M NaCl medium) was used to characterize the stoichiometry and stability of the species formed in the interactions of the Al(III) ion and the peptide GSSG, while multinuclear ((1)H, (13)C, (27)Al) nuclear magnetic resonance (NMR) and electrospray mass spectroscopy (ESI-MS) were applied to characterize the binding sites and species of the metal ion in the complexes. Two-dimensional ((1)H, (1)H-NOESY) was also employed to reveal the difference in the conformational behavior of the peptide and its complexes. The following results were obtained: (1) Aluminum(III) can coordinate with the important biomolecule GSSG through the following binding sites: glycyl and glutamyl carboxyl groups to form various mononuclear 1:1 (AlLH(4), AlLH(3), AlLH(2), AlLH, AlL, AlLH(-1), AlLH(-2)) and several binuclear 2:1 (Al(2)LH(4), Al(2)LH(2), Al(2)L) species (where H(6)L(2+) denotes the totally protonated oxidized glutathione) in acidic aqueous solutions. (2) It indicates that the COO(-) groups at low level of preorganization in such small peptide are not sufficient to keep the Al(III) ion in solution and to prevent the precipitation of Al(OH)(3) in the physiological pH range. (3) It also suggests that the occurrence of an Al-linked complexation, the conformation of the peptide GSSG in aqueous solutions appeared to change a little, relative to the initial structure.

[Study on the effect of aluminum (III) on NADH in the presence of NH4Ac in aqueous solution].

This paper studied the interactions of Al (III ) and dihydronicotinamide adenine dinucleotide (NADH) in nearly neutral aqueous solutions (pH 6.5) by means of UV-Vis and 1H, 13C-NMR techniques. The results suggested that Al (III) interacts with NADH to form Al-NADH complexes by occupying the binding sites of phosphate oxygen atoms O(N)1 and O(A)1 and ribose ring hydroxyl groups, which are the potential recognition sites for substrates, coenzyme and enzyme. In the presence of NH4Ac salt buffer and with Al (III) salt solution, NADH will be marked with structural changes at the nicotinamide moiety in contrast with almost no structural changes in Tris-HCl buffer solution with Al (III) salt.