OBJECTIVE To design and synthesize mono-carbonyl curcumin analogues（MCACs） and investigate the activities of them against breast cancer. METHODS The analogues F1， F2， and F3 were obtained by aldol condensation reaction， and their antitumor activities（including the activities of human breast cancer cell MCF-7 and human lung cancer cell A549） were detected by MTT assay [evaluated with half inhibitory concentration（IC50）]. The results of MTT assay were compared with those of curcumin. Bioinformatics methods were used to collect the core targets of analogues F1， F2 and F3 acting on breast cancer， and then molecular docking verification was carried out. The cell experiments were conducted to investigate the effects of high， medium and low concentrations （16， 8， 4 μmol/L） of F1， F2 and F3 on the expression of the first core target protein as well as the effects of medium concentration of F1， F2 and F3 on the expression of cleaved-caspase-3. RESULTS Compared with curcumin， IC50 of analogues F1， F2 and F3 to A549 and MCF-7 cells（except for IC50 of analogue F2 to A549 cells） were decreased significantly（P＜ 0.05 or P＜0.01）； among them， IC50 of analogue F2 to MCF-7 cell was the lowest， being（9.67±1.27） μmol/L. Bioinformatics analysis showed that index of affinity of analogues F1， F2 and F3 with the first core target epidermal growth factor receptor （EGFR）， protein kinase B （AKT） and AKT were 5.909 2， 8.402 5 and 6.486 6， respectively； high concentration of F1 could significantly reduce the phosphorylation level of EGFR protein in MCF-7 cells（P＜0.01）， while low， medium， and high concentrations of F2 and high concentration of F3 could significantly reduce the phosphorylation level of AKT protein in MCF-7 cells（P＜0.05 or P＜0.01）. Medium concentration of F1， F2， and F3 could significantly increase the expression level of cleaved- caspase-3 protein in MCF-7 cells（P＜0.01）. CONCLUSIONS Designed and synthesized MCACs F1， F2 and F3 all have good anti- breast cancer activity， and F2 has better anti-breast cancer activity.

Objective To investigate the effect and mechanism of a high uric acid environment on cataract formation by rat lens ex-vivo culture model.Methods The lenses harvested from adult Sprague Dawley rats were cultured in a high uric acid environment in vitro.The lenses were divided into the blank control group(15 eyes)and the uric acid treatment group(15 eyes).The lenses in the uric acid treatment group were treated with M-199 medium with 800 μmol·L-1 uric acid and lenses in the blank control group were cultured with M-199 medium without uric acid for 7 days.The turbidity of the lenses of rats in the two groups was observed by optical microscope.The deposition of urate in the lenses of rats in the two groups was detected by the Gomori hexamine silver method.The senescence of lens cells in the two groups was measured by SA-[3-gal staining.Lens cell apoptosis in the two groups was detected by the TUNEL method.Results The opacifica-tion in lenses of the uric acid treatment group occurred on the 5th day and the opacification grade significantly increased on the 7th day,with opacification grades being significantly different from those in the blank control group(both P＜0.05).In the blank control group,the lens epithelial cells were arranged neatly with no urate deposition,a few blue-stained lens epi-thelial cells and no obvious positive cells.In the uric acid treatment group,the arrangement of the lens epithelial was disor-dered in the equatorial part of the lens and under the anterior capsule,with black-brown urate crystals around the lens,more blue-stained lens epithelial cells and a small number of positive cells.In the high uric acid environment,urate crystals deposited under the capsular membrane of the lens,the surrounding lens epithelial cells were senescent,and a small num-ber of lens epithelial cells died.Conclusion A high uric acid environment can steadily and effectively induce cataract formation in the rat lens ex-vivo culture model.The mechanism may be related to senescence and apoptosis of lens epitheli-al cells induced by uric acid.

Objective:To investigate the specific method, efficacy and safety of intrathecal baclofen (ITB) in the treatment of moderate and severe spasticity.Methods:The clinical features, response to ITB trial, clinical outcomes and treatment-related complications after baclofen pump implantation surgery were retrospectively analyzed in patients with moderate to severe spasticity who received ITB treatment in Hong Kong University Shenzhen Hospital from August 2022 to August 2023.Results:The study included 22 patients with moderate to severe spasticity, of whom 18 patients (81.8%) were effective in ITB trial. For these patients with effective trial, the lowest mean Modified Ashworth Scale (MAS) score of lower extremity muscle groups after ITB bolus injection was decreased significantly compared with the baseline mean value before injection (1.29±0.81 vs 3.31±1.18 , t=11.364, P<0.001), and the M/H amplitude ratio of soleus H-reflex at 4 hours after ITB bolus injection was significantly higher than the baseline value (9.61±4.66 vs 3.83±2.69, t=7.780, P<0.001). Among the 18 patients with effective ITB trial, 10 patients received baclofen pump implantation surgery with the informed consent of the patients or their guardians. Compared with the 8 patients without surgery, the baseline mean MAS score of lower extremity muscle groups of the 10 patients who received surgery was higher (3.71±1.06 vs 2.83±1.16, F=5.645, P<0.05), the maximum reduction of mean MAS score of lower extremity muscle groups of the 10 patients who received surgery was greater (2.61±0.99 vs 1.30±0.64, F=20.696, P<0.001). In 10 patients who received baclofen pump implantation surgery, the mean MAS score of lower limb muscle groups was significantly decreased in the first and sixth months after surgery compared with the baseline mean value before surgery (1.64±0.91, 1.16±0.89 and 3.71±1.06, F=39.946, P<0.001). Among them, 2 patients were followed up to 12 months after surgery, 2 patients were followed up to 18 months after surgery, and the mean MAS score of their lower limb muscle groups showed continuous improvement, and 1 patient with cerebral palsy combined with lower urinary tract hyperactive syndrome also showed significant improvement of urgent urinary incontinence after surgery. Up to the last follow-up, the daily dose of baclofen in 10 patients receiving baclofen pump implantation was (143.60±96.24) μg, and no treatment-related complications occurred. Conclusions:ITB can effectively relieve spasticity symptoms and other related neurological dysfunction in patients with moderate and severe spasticity, and the efficacy can be maintained for a long time, and treatment-related complications are rare. Preoperative ITB trial is helpful to screen out patients more suitable for baclofen pump implantation. Patients with higher grade of spasticity at baseline and better efficacy of ITB trial are more suitable for baclofen pump implantation. The soleus H reflex can effectively reflect the change of stretch reflex of lower limb muscle groups in patients with spasticity treated by ITB, and can be used to evaluate the treatment effect of ITB.

OBJECTIVE To establish a quantitative analysis of multi-components by single marker （QAMS） method based on a variety of internal reference substances for the content determination of 6 components in Jinlian qingre granules， such as mangiferin， 2″-O-β-L-galactopyranosylorientin， orientin， veratric acid， vitexin， harpagoside. METHODS The determination was performed on Agilent Eclipse Plus C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at the flow rate of 1 mL/min. The column temperature was 30 ℃， and the detection wavelength was set at 270 nm. Taking orientin， vitexin and 2″-O-β-L-galactopyranosylorientin as internal references， the relative correction factors （RCF） of the other 5 components to be determined and internal substances were determined by QAMS. The contents of 6 components in 21 batches of Jinlian qingre granules were calculated and then compared with the results of the external standard method. RESULTS The contents of mangiferin， 2″-O-β-L-galactopyranosylorientin， orientin， veratric acid， vitexin and harpagoside in 21 batches of samples were determined by QAMS in the range of 0.234-0.516， 1.804-2.270， 2.143-2.606， 0.190-0.223， 0.594-0.782， 0.080-0.152 mg/g； the contents of them determined by external standard method were 0.235-0.523， 1.798-2.265， 2.137-2.599， 0.190-0.224， 0.597-0.786， 0.077-0.151 mg/g， respectively. The percentage difference between the results measured by the two methods should not exceed 4.00%. CONCLUSIONS QAMS has been constructed for the simultaneous determination of 6 components in Jinlian qingre granules based on a variety of internal reference substances. The results obtained by this method are not significantly different from those obtained by the external standard method， and can be used for the quality control of Jinlian qingre granules.

Humans ; Sodium-Calcium Exchanger ; Carrier Proteins ; Pain ; Calcium/metabolism*

Humans ; Glioblastoma ; Cell Line, Tumor ; Neoplasm Invasiveness ; Cell Movement ; Brain Neoplasms

Objective:To investigate the effect of epigallocatechin gallate (EGCG) on the activation of human Tenon fibroblasts (HTFs) and its mechanism.Methods:Tenon capsule tissues from nine eyes of nine advanced primary open angle glaucoma patients during trabeculectomy were obtained for primary cell culture.HTFs harvested were identified by immunofluorescence staining for vimentin and keratin.Cells at passage 4-6 were used for experiment.Viability of HTFs treated with EGCG at 0, 10, 20, 30, 40, 50, 60, 70 and 80 μmol/L was detected by cell counting kit-8 (CCK-8) assay.The cells were divided into blank control group, transforming growth factor (TGF)-β1-induced group, and EGCG-treated group, which were cultured in normal medium, medium containing 10 ng/ml TGF-β, medium containing 10 ng/ml TGF-β+ 50 μmol/L EGCG, respectively.The proliferation rate of HTFs was detected by BrdU labeling assay.Cell migration was observed by scratch wound healing assay.The expression of α-smooth muscle actin (α-SMA) was measured by immunofluorescence staining.The protein relative expression levels of Smad2/3, phosphoinositide-3-kinase (PI3K), protein kinase B (Akt) as well as the phosphorylated Smad2/3 (p-Smad2/3) and phosphorylated Akt (p-Akt) were measured by western blot.This study was approved by the Ethics Committee of Guangdong Provincial People's Hospital (NO.GDREC2019331H[R1]).Results:The HTFs harvested had spindle shape, grew regularly and were vimentin-positive.CCK-8 assay showed that there was no significant difference in the variability of HTFs treated with EGCG at 10, 20, 30, 40 and 50 μmol/L in comparison with 0 μmol/L EGCG treatment (all at P<0.05). BrdU labeling assay showed that cell proliferation in the TGF-β1-induced group was (66.37±12.65)%, which was significantly higher than (14.75±12.33)% in EGCG-treated group ( P<0.05). Three days after scratch, the relative scratch area in the TGF-β1-induced group was (47.33±12.22)%, which was significantly lower than (92.67±4.04)% in the EGCG-treated group ( P<0.05). Immunofluorescence assay showed that α-SMA fluorescence was significantly enhanced in the TGF-β1-induced group in comparison with the blank control group, which was reduced to blank control group level in EGCG-treated group.Western blot analysis showed that there were significant differences in the relative expression levels of p-Smad2/3, PI3K and p-Akt protein among the various groups ( F=58.820, 121.153, 69.289; all at P<0.001). The relative expressions of p-Smad2/3, PI3K and p-Akt in the TGF-β1-induced group were significantly higher than those in the blank control group, 10 μmol/L and 50 μmol/L EGCG-treated groups (all at P<0.05). Conclusions:EGCG can suppress TGF-β1-induced HTFs activation through Smad and PI3K/Akt signaling pathways.

Nanotechnology has emerged as an ideal approach for achieving the efficient chemo agent delivery. However, the potential toxicity and unclear internal metabolism of most nano-carriers was still a major obstacle for the clinical application. Herein, a novel "core‒shell" co-assembly carrier-free nanosystem was constructed based on natural sources of ursolic acid (UA) and polyphenol (EGCG) with the EpCAM-aptamer modification for hepatocellular carcinoma (HCC) synergistic treatment. As the nature products derived from food-plant, UA and EGCG had good anticancer activities and low toxicity. With the simple and "green" method, the nanodrugs had the advantages of good stability, pH-responsive and strong penetration of tumor tissues, which was expected to increase tumor cellular uptake, long circulation and effectively avoid the potential defects of traditional carriers. The nanocomplex exhibited the low cytotoxicity in the normal cells

Objective:To investigate the clinical effect of transsphincter fistulectomy in the treatment of anal fistula.Methods:Seventy-three patients with anal fistula who received treatment in Zhoushan Hospital from March 2016 to March 2020 were included in this study. They were randomly assigned to undergo either conventional incision combined with thread-drawing drainage (control group, n = 35) or transsphincter fistulectomy (observation group, n = 38). Operative time, wound healing time, length of hospital stay, Visual Analogue Scale (VAS) score 24 and 48 hours after surgery, complications, the improvement in anal sphincter function before and 3 months after surgery were compared between the two groups. Results:Operative time, wound healing time and length of hospital stay in the observation group were (49.83 ± 7.67) minutes, (20.78 ± 3.54) days and (5.31 ± 1.27) days, which were significantly shorter than those in the control group [(62.31 ± 5.45) minutes, (25.87 ± 3.10) days, (7.78 ± 1.32) days, t = 8.063, 6.512, 8.133, all P < 0.05). The VAS score 24 and 48 hours after surgery in the observation group were (2.43 ± 0.64) points and (1.21 ± 0.36) points, respectively, which were significantly lower than those in the control group [(3.87 ± 1.23) points, (2.83 ± 0.97) points, t = 6.347 and 9.607, both P < 0.05]. The incidence of complication in the observation group was significantly lower than that in the control group [5.26% (2/38) vs. 28.57% (10/35), χ2 = 7.206, P < 0.05]. Conclusion:Transsphincter fistulectomy in the treatment of anal fistula has good therapeutic effects, can reduce pain and has little impact on the function of anal sphincter. It is innovative and scientific.

Objective:To investigate the clinical effect of local and systemic zoledronic acid in the treatment of giant cell tumor of bone.Methods:The clinical data of 42 patients with giant cell tumor of bone who were treated in the Department of Joint and Sports Medicine Surgery of Shandong Provincial Third Hospital from January 2000 to January 2017 were retrospectively analyzed. According to whether zoledronic acid was used during and after operation, the patients were divided into zoledronic acid group ( n=21) and non zoledronic acid group ( n=21). The perioperative indexes, pain visual analogue scale (VAS), international Musculoskeletal Tumor Society (MSTS) score of lower extremity function, adverse reactions and the postoperative recurrence were compared between the two groups. Results:The operative time of zoledronic acid group and non zoledronic acid group were (158.4±20.5) min and (169.5±19.5) min, the intraoperative bleeding volume were (236.3±9.7) ml and (228.2±16.5) ml, the postoperative drainage volume were (163.3±7.4) ml and (161.4±9.3) ml, and the healing time of incision were (13.8±2.1) d and (14.0±2.0) d, respectively, with no significant difference ( t=-1.798, P=0.080; t=1.936, P=0.062; t=0.733, P=0.468; t=-0.290, P=0.774). The preoperative VAS scores of zoledronic acid group and non zoledronic acid group were 6.54±1.76 and 6.72±1.51 respectively, the MSTS scores were 13.56±2.35 and 12.79±1.98 respectively, and the differences were not statistically significant ( t=-0.356, P=0.724; t=1.148, P=0.258). The VAS scores of the two groups were 1.32±0.31 and 1.92±0.19 at 4 weeks after operation, 0.93±0.29 and 1.47±0.38 at 3 months after operation respectively, and the differences were statistically significant ( t=-7.562, P<0.001; t=-5.177, P<0.001). The VAS scores of the two groups were 0.31±0.12 and 0.35±0.23 at the last follow-up, with no significant difference ( t=0.707, P=0.485). The MSTS scores of zoledronic acid group and non zoledronic acid group were 24.89±3.86 and 21.82±2.95 at 4 weeks after operation, 26.78±2.57 and 24.62±2.62 at 3 months after operation respectively, and the differences were statistically significant ( t=2.896, P=0.006; t=2.697, P=0.010). The MSTS scores of the two groups were 27.31±2.21 and 26.69±2.93 at the last follow-up, with no significant difference ( t=0.774, P=0.443). The postoperative recurrence time of the two groups was (9.79±2.58) months and (7.31±1.73) months respectively, and the difference was statistically significant ( t=3.659, P=0.001). There was no significant difference in recurrence Campanacci grade and recurrence tumor location between the two groups ( U=7.000, P=0.860; χ2=1.062, P>0.999). The occurrence rates of fever in zoledronic acid group and non zoledronic acid group were 23.81% (5/21) and 4.76% (1/21), the occurrence rates of myalgia were 19.05% (4/21) and 4.76% (1/21), the incidences of influenza like symptoms were 14.29% (3/21) and 0 (0/21), the occurrence rates of gastrointestinal reaction were 9.52% (2/21) and 4.76 (1/21), and the differences were not statistically significant ( χ2=1.750, P=0.186; χ2=0.980, P=0.341; χ2=1.436, P=0.231; χ2<0.001, P>0.999). All the patients had no serious adverse reactions such as liver and kidney function damage and mandible necrosis. Conclusion:Local and systemic application of zoledronic acid in the treatment of giant cell tumor of bone can improve the early postoperative pain and limb function, delay the recurrence time, and can be used as an auxiliary treatment of giant cell tumor of bone.