[Topography-guided transepithelial corneal collagen cross-linking by sequential ultraviolet A irradiation in different diameters for progressive keratoconus in adults].

Objective: To compare the efficacy and safety of a novel customized topography-guided transepithelial corneal collagen cross-linking (TG-CXL) procedure by sequential ultraviolet A irradiation in different diameters and conventional transepithelial corneal collagen cross-linking (TE-CXL) in adult patients with progressive keratoconus. Methods: A prospective cohort study was conducted. Adult patients diagnosed with progressive keratoconus in the Affiliated Xiamen Eye Center of Xiamen University were continuously recruited and randomly assigned to receive the TG-CXL or TE-CXL procedure from March 2020 to March 2021. Patients in the TE-CXL group were irradiated in the central 9-mm zone of the cornea (total energy, 7.2 J/cm2; irradiance, 45 mW/cm2), while patients in the TG-CXL group were first irradiated with the protocol used in the TE-CXL group, and further irradiated in the central 6-mm zone (total energy, 3.6 J/cm2; irradiance, 9 mW/cm2). The subjective symptom of pain and corneal fluorescein sodium staining were scored within postoperative 3 days. Slit lamp examination, measurements of uncorrected visual acuity (UCVA) and best-corrected visual acuity (BCVA), corneal topography, anterior segment optical coherence tomography, in vivo corneal confocal microscopy, corneal endothelial cell count, and non-contact tonometry were performed before surgery and at 3, 6, and 12 months after surgery. Results: A total of 66 patients were enrolled (mean age, 23.0±3.3 years old), with 33 patients (33 eyes) in each group. No statistically significant differences were found in age, gender, and maximum keratometry (Kmax) between the two groups (P>0.05). On day 1 after surgery, the average pain score of the TG-CXL group (2.21±0.45) was significantly higher than that of the TE-CXL group (1.32±0.33) (P<0.05). The pain was rapidly alleviated in both groups on days 2 and 3. On days 1 and 2, the corneal fluorescein sodium staining scores in the TG-CXL group (4.15±0.83 and 2.21±0.60, respectively) were significantly higher than those in the TE-CXL group (1.76±0.56 and 0.85±0.51, respectively, P<0.001), while there was no significant difference between the two groups at day3 (P=0.184). The UCVA and BCVA of the TG-CXL group at 3, 6, and 12 months after surgery were significantly improved when compared with the baseline. At 3, 6, and 12 months, the BCVA (LogMAR) of the TG-CXL group (0.21±0.15, 0.22±0.16, and 0.22±0.16, respectively) were significantly improved when compared with those of the TE-CXL group(0.32±0.15, 0.34±0.15, and 0.36±0.16, respectively, P<0.01). However, there was no significant difference in UCVA between groups at any time point after surgery (P>0.05). The spherical and cylindrical power values of the TG-CXL group were improved when compared with the baseline (P<0.05). However, no significant difference in spherical power values was found between the two groups at any time point after surgery (P>0.05). Meanwhile, there were significant differences in cylindrical power values between the two groups at 6 and 12 months after surgery (P<0.05). The Kmax in the TG-CXL group was improved at all of the time points after surgery when compared with the baseline (P<0.001), while no significant difference in Kmax was found at any time point after surgery in the TE-CXL group when compared with the baseline (P>0.05). At 6 and 12 months after surgery, the Kmax values in the TG-CXL group were significantly lower than the TE-CXL group (P<0.05). No significant differences were found in flat keratomety, steep keratometry, the minimal thickness of the cornea, endothelial cell density, and intraocular pressure between the two groups at any time point after surgery (P>0.05). Within one month after surgery, optical coherence tomography revealed the increased density in the anterior stroma in both groups. In most patients in the TG-CXL group, a demarcation line was visible in the central and para-central corneal stroma, representing a clear and continuous, high-signal arc-shaped linear structure, which was deeper in the central cornea than the para-central cornea. In contrast, a demarcation line, fuzzy and focally discontinuous, was visible only in a few patients in the TE-CXL group, with an almost uniform depth in the central and the para-central cornea. Confocal microscopy demonstrated an apparent mesh-like cross-linked collagen structure in the superficial and intermediate corneal stroma at all time points after surgery in the TG-CXL group, with thickening stromal collagen fibers and an increased number of interconnections. In contrast, the mesh-like structure and number of interconnections in the superficial corneal stroma were significantly reduced at 12 months after surgery in the TE-CXL group, with no cross-linking structure in the intermediate corneal stroma at any time point after surgery. No serious complications such as corneal infection, sterile corneal ulcer, and persistent epithelial defect were observed in both groups during the follow-up of 12 months. Conclusions: The TG-CXL procedure by sequential irradiation in two different diameters with ultraviolet A light was effective and safe in the management of progressive keratoconus in adults, achieving significant refractive improvement. This might be a good technical alternative for refractive corneal cross-linking surgery.

[Efficacy of deproteinized calf blood extract eye drops on early recovery after pterygium surgery].

Objective: To investigate the effect of deproteinized calf blood extract eye drops on early postoperative recovery in primary pterygium patients. Methods: This is a prospective randomized controlled study. Patients diagnosed with primary pterygium in single eye at affiliated Xiamen Eye Center of Xiamen University during March 2016 to May 2016 were enrolled. After Pterygium excision with autologous conjunctival transplantation, patients were randomly assigned into four groups by a random number table, treated with anti-inflammaroty drugs only (control group) or combined with the following agents: deproteinized calf blood extract eye drops (DCBE group), carboxymethylcellulose sodium eye drops (CMC group), and recombinant human epidermal growth factor eye drops (rEGF group). Short-form McGill pain questionnaire, slit lamp and corneal fluorescein sodium staining, non-contact intraocular pressure, uncorrected visual acuity (UCVA) and best corrected visual acquity (BCVA) as well as redness score of bulbar conjunctiva were performed before surgery (d0) and on day 1 (d1), day 2 (d2), day 3 (d3), day 7 (d7) and day 14 (d14) after surgery. Results: One hundred and fourteen patients including 43 males and 71 females, aged (48.9±12.5) years, were eventually included in this study. The McGill scores gradually decreased after surgery in all groups. On d2, the McGill score in DCBE group, control group, CMC group and rEGF group was (1.42±0.67), (2.21±0.88), (1.93±1.08) and (1.77±1.18), respectively; On d3, the score was (1.32±0.54), (1.93±0.72), (1.79±0.87) and (1.52±0.77), respectively. On d2 and d3, statistical difference was recorded among groups (d2, F=3.43, P=0.019; d3, F=4.047, P=0.009), and the McGill score of DCBE group was significantly lower than that of CMC group (d2, P=0.047, d3, P=0.017). On d2, the percentage of corneal epithelium defect in DCBE group, control group, CMC group and rEGF group was 8.6%±1.9%, 11.7%±1.7%, 11.5%±1.9% and 10.4%±1.8%, respectively; On d3, the percentage was 4.5%±2.2%, 9.2%±2.4%, 7.4%±2.5% and 5.9%±2.3%, respectively. On d2 and d3, statistical difference of corneal epithelium defect percentage was recorded among groups (d2, F=17.17, P<0.001; d3, F=21.4, P<0.001). On d2, the percentage of corneal epithelium defect in DCBE group was significantly lower than the other three groups (P<0.01); On d3, the percentage of corneal epithelium defect in DCBE group was significantly lower than control group and CMC group (P<0.001), while no difference was found between DCBE group and rEGF group (P>0.05). However, no statistical differences were recorded in the number of patients with vision improvement among the groups (P>0.05). The intraocular pressure remained stable. No differences in the conjunctival redness score were found among the groups after surgery (P>0.05). Conclusion: Our data demonstrated the efficacy of deproteinized calf blood extract eye drops on the postoperative management in patients with primary patients, with faster pain relief and promoted epithelium recovery. (Chin J Ophthalmol, 2019, 55:134-140).

[Accelerated transepithelial corneal collagen cross-linking for progressive keratoconus with a thin cornea: one-year results].

Objective: To evaluate the clinical results of keratoconic eyes with a thin cornea treated with accelerated transepithelial corneal collagen cross-linking (A-TE-CXL) within 1 year. Methods: Nineteen eyes of 19 patients with progressive keratoconus with a minimum corneal thickness from 380 µm to 420 µm (including the epithelium) were included in this prospective, nonrandomized clinical study and treated with A-TE-CXL. Scoring of pain and foreign body sensation, slit lamp examination, uncorrected visual acuity, best corrected distance visual acuity, corneal topography, anterior segment optical coherence tomography, in vivo corneal confocal microscopy and endothelial cell count were assessed before surgery and at 1, 3, 6 and 12 months postoperatively. Paired t test was applied for statistical analysis. Results: Mild pain and moderate foreign body sensation were reported by most patients within postoperative 24 hours, but rapidly disappeared on day 2. Extremely mild epithelial damage was observed within postoperative 24 hours, and the epithelium fully recovered on day 2. Improvement of visual acuity was recorded at 3 and 12 months. Pentacam corneal topography revealed a significant reduction of the thickness of the thinnest location from(395.2±13.8)µm preoperatively to (378.9±17.1)µm at 1 month postoperatively (t=2.982, P<0.01). Front curvature values were reduced postoperatively. K(MAX) was significantly decreased at 12 months (55.67±4.91) compared with (57.35±5.54) preoperatively, while K2 was also significantly decreased at 12 months (52.18±3.70) compared with (52.70±3.56) preoperatively (K(MAX), t=3.044, P<0.01. K2, t=2.384, P<0.05) . Within 1 month postoperatively, optical coherence tomography exhibited an increase of reflectance with a demarcation line in the anterior stroma. In vivo confocal microscopy also showed significant thickening and increased connections of collagen fibers with a maximal depth at about 90 to 120 µm. The corneal endothelial cell density remained stable (t=0.692, P>0.05). None of the patients showed postoperative complications such as corneal infection, scarring and ulceration. Conclusions: Within 1 year postoperatively, A-TE-CXL was effective and safe for the management of progressive keratoconus with a thin cornea. A-TE-CXL showed the advantages of very short time consuming in surgery, rapid recovery and very few complications, and had the potential to become a valid alternative for the treatment of keratoconus. (Chin J Ophthalmol, 2017, 53: 694-700).

[Study on lignans from Diphylleia sinensis].

A new lignan along with eight known lignans and a flavonoid were isolated from the rhizomes of Diphylleia sinensis Li.. Their structures were elucidated on the basis of chemical and spectral analysis. The new lignan, compound IX, was named as picropodophyllin-1-ethyl ether (IX). The eight known lignans were identified as podophyllotoxin (I), isopicropodophyllone (II), dehydropodophyllotoxin (III), diphyllin (IV), picropodophyllin (V), podophyllotoxone (VI), 4'-demethylpodophyllotoxin (VII) and picropodophyllin glucoside (VIII). Compounds II, VI, VII and VIII were found for the first time from the rhizome of this plant. The flavonoid was identified as kaempferol.

[Determination of gelsemium alkaloids by RP-HPLC].

Gelsemium elegans Benth is a kind of traditional Chinese medicine, eight alkaloids have been isolated from this herb. In recent years, an HPLC method for the separation and determination of five of these alkaloids, ie. gelsemium A (G), koumine (F), kumantenidine (D), kumantenine (B) and kumantenmine (A) is described, dichroine being used as the internal standard. In this report a RP column of C18 and the mobile phase methanol--water--n-butylamine (78:22:0.1 V/V) were employed. The flow rate was 1.0 ml/min, the column temperature was 25 degrees C and the detection wavelength was 256 nm. The calibration curves showed good linearity over the range of 0.02-0.12 microgram, r = 0.9835-0.9977 and the recoveries were 95.01-99.70% for the five alkaloids. The method is simple, sensitive and reproducible and can be used for the quality control of Gelsemium preparations for clinical evaluation.

[Studies on the TLC scanning determination of lignans in Diphylleia sinensis Li].

A simple, sensitive and accurate method for the separation and determination of the lignans: podophylltoxone (I), isopicropodophyllone (II), picropodophyllone (III), dehydropodophyllotoxin (IV), picropodophyllin (V), podophyllotoxin (VI), 4'-demethylpodophyllotoxin (VII) and diphyllin (VIII) is described. The sample solution was applied at a point 1 cm from the bottom edge of the HPTLC silica gel plate (10 cm x 10 cm), dichloromethane-diethyl ether (4:1) was used as the developing solvent. The plate was saturated for 30 min and then developed twice for 9.5 cm using ascending technique. The plate was sprayed with 2.5% ammonium ceric sulphate--20% nitric acid and toasted for 15 min at 120 degrees C, then fumigated with ammonia solution for 20 min at room temperature to intensify the spot color. The spots were scanned with a Shimadzu CS-930 TLC scanner. The contents of eight lignans in Diphylleia sinensis was calculated by comparison with standards spotted on the same plate. The standard curves were linear in the range of 0.48-2.52 micrograms for the eight lignans. The method has been applied to the analysis of various samples and can be used for the quality control of Diphylleia sinensis, podophyllum and dysosma preparations used in clinic.

[Comparative study of the relationship between in vitro motility and invasion potentials of three mouse forestomach cancer (MFC) cell lines].

The relationship between motility of cancer cells and their invasiveness is important in understanding the invasion mechanisms of malignant tumors. The in vitro motility of three MFC cell lines was measured by the Boyden chamber technique. Statistical analysis of the results showed significant differences in vitro motility among these three cell lines. The motility correlated with in vitro invasion potentials.

[Determination of naphthoquinones in tissue cultured Zicao].

A simple, sensitive and accurate method for the separation and determination of the naphthoquinones, deoxyshikonin (I), acetylshikonin (II) and beta-beta-dimethylacrylshikonin (III) in tissue cultured Zicao is described. The sample solution was applied at a point 1 cm from the bottom edge of the TLC silica gel plate (10 x 15 cm). A mixture of n-hexane-acetone-chloroform-glacial acetic acid (13:0.3:0.2:0.05 v/v) was used as the developing solvent. The plate was saturated for 30 min and then developed twice for 15 cm using ascending technique. The spots were scraped off and extracted with CHCl3, then determined spectrophotometrically. The standard curve was linear in the range of 0.65-2.25 micrograms for the naphthoquinones. The method has been applied to the analysis of different samples. This method can be used for the quality control of naphthoquinones in tissue culture products.

Establishment, characteristics, and utilization of a new in vivo-in vitro system.

Radiotherapy and chemotherapy are two important methods for malignant tumor treatment. To research radiobiological response in therapy, we have established a better experimental method in contrast to the traditional ones such as TCD50, regrowth delay, cell survival curve, etc, all with their limitations. A new mouse tumor in vivo-in vitro system LA795 Vv-Vt has been developed for studies on radiobiology. Such a system could be used to study the in vivo response of a solid tumor by the in vitro cloning assay. For the purpose of increasing the PE in vitro, LA795 Vv-Vt tumor line was purified through culturing the cells as a clonogenic spheroid. The spheroids were then injected into the flank of mouse subcutaneously for tumor growth. The in vivo-in vitro system LA795 Vv-Vt is an excellent model dissecting and analyzing the various factors which affect tumor development and determine the response of tumor to specific agent and regimens.