[Antitumor effects of redox-responsive nanoparticles containing platinum（⠣）in ovarian cancer].

Objectives: To explore the antitumor effects of redox-responsive nanoparticles containing platinum(Ⅳ)-NP@Pt(Ⅳ) in ovarian cancer. Methods: Redox-responsive polymer carriers were synthesized. Polymer carriers and platinum(Ⅳ)-Pt(Ⅳ) can self-assemble into NP@Pt(Ⅳ). Inductively coupled plasma mass spectrometry was performed to detect the platinum release from NP@Pt(Ⅳ) in reducing environment and the platinum content in ovarian cancer cells ES2 treated with cisplatin, Pt(Ⅳ) and NP@Pt(Ⅳ). The proliferation ability of the ovarian cancer cells were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cellular apoptosis was assessed by flow cytometry. Collection of primary ovarian cancer tissues from patients with primary high-grade serous ovarian cancer who were surgically treated at the Cancer Hospital of the Chinese Academy of Medical Sciences from October to December 2022. The high-grade serous ovarian cancer patient-derived xenograft (PDX) mice were intravenously injected with Cy7.5 labeled NP@Pt(Ⅳ) followed by in vivo imaging system. Mice were treated with PBS, cisplatin and NP@Pt(Ⅳ). Tumor volume and weight were measured in each group. Necrosis, apoptosis and cell proliferation of tumor tissues were detected by hematoxylin-eosin (HE) staining, TUNEL fluorescence staining and Ki-67 immunohistochemistry staining. Body weight and HE staining of heart, liver, spleen, lung and kidney of mice in each group were measured. Results: The platinum release of NP@Pt(Ⅳ) after 48 hours in reducing environment was 76.29%, which was significantly higher than that of 26.82% in non-reducing environment (P<0.001). The platinum content in ES2 cells after 4 hours and 7 hours of treatment with NP@Pt(Ⅳ) (308.59, 553.15 ng/million cells) were significantly higher than those of Pt(Ⅳ) (100.21, 180.31 ng/million cells) and cisplatin (43.36, 50.36 ng/million cells, P＜0.05). The half inhibitory concentrations of NP@Pt(Ⅳ) in ovarian cancer cells ES2, A2780, A2780DDP were 1.39, 1.42 and 4.62 µmol/L, respectively, which were lower than those of Pt(IV) (2.89, 7.27, and 16.74 µmol/L) and cisplatin (5.21, 11.85, and 71.98 µmol/L). The apoptosis rate of ES2 cells treated with NP@Pt(Ⅳ) was (33.91±3.80)%, which was significantly higher than that of Pt(Ⅳ) [(16.28±2.41)%] and cisplatin [(15.01±1.17)%, P＜0.05]. In high-grade serous ovarian cancer PDX model, targeted accumulation of Cy7.5 labeled NP@Pt(Ⅳ) at tumor tissue could be observed. After the treatment, the tumor volume of mice in NP@Pt(IV) group was (130±98) mm3, which was significantly lower than those in control group [(1 349±161) mm3, P<0.001] and cisplatin group [(715±293) mm3, P=0.026]. The tumor weight of mice in NP@Pt(IV) group was (0.17±0.09)g, which was significantly lower than those in control group [(1.55±0.11)g, P<0.001] and cisplatin group [(0.82±0.38)g, P=0.029]. The areas of tumor necrosis and apoptosis in mice treated with NP@Pt(Ⅳ) were higher than those in mice treated with cisplatin. Immunohistochemical staining revealed that there were low expressions of Ki-67 at tumor tissues of mice treated with NP@Pt(Ⅳ) compared with cisplatin. The change in body weight of mice in NP@Pt(Ⅳ) group was not significantly different from that of the control group [(18.56±2.04)g vs.(20.87±0.79)g, P=0.063]. Moreover, the major organs of the heart, liver, spleen, lung, and kidney were also normal by HE staining. Conclusion: Redox-responsive NP@Pt(Ⅳ), produced in this study can enhance the accumulation of cisplatin in ovarian cancer cells and improve the efficacy of ovarian cancer chemotherapy.

[Signal transducer and activator of transcription 3 and cancer associated fibroblasts jointly generate chemo-resistance and affect prognosis in epithelial ovarian cancer].

Objective: To investigate the mechanism of signal transducer and activator of transcription 3 (STAT3) and cancer associated fibroblasts (CAF) jointly generate chemo-resistance in epithelial-ovarian cancer and their effect on prognosis. Methods: A total of 119 patients with high-grade ovarian serous cancer who received surgery in Cancer Hospital of Chinese Academy of Medical Sciences from September 2009 to October 2017 were collected. The clinico-pathological data and follow-up data were complete. Multivariate Cox regression model was used to analyze the prognostic factors. Ovarian cancer tissue chips of patients in our hospital were prepared. EnVision two-step method immunohistochemistry was used to detect the protein expression levels of STAT3, the specific markers of CAF activation, fibroblast activating protein (FAP), and type Ⅰ collagen (COL1A1) secreted by CAF. The relationship between the expression of STAT3, FAP, COL1A1 protein and drug resistance and prognosis of ovarian cancer patients was analyzed, and the correlation between the expression of three proteins was analyzed. These results were verified through the gene expression and prognostic information of human ovarian cancer tissues collected in the GSE26712 dataset of gene expression omnibus (GEO) database. Results: (1) Multivariate Cox regression model analysis showed that chemotherapy resistance was an independent risk factor for overall survival (OS) of ovarian cancer (P<0.001). (2) The expression levels of STAT3, FAP, and COL1A1 proteins in chemotherapy resistant patients were significantly higher than those in chemotherapy sensitive patients (all P<0.05). Patients with high expression of STAT3, FAP, and COL1A1 had significantly shorter OS than those with low expression (all P<0.05). According to the human ovarian cancer GSE26712 dataset of GEO database, patients with high expression of STAT3, FAP, and COL1A1 also showed shorter OS than patients with low expression (all P<0.05), the verification results were consistent with the detection results of ovarian cancer patients in our hospital. (3) Correlation analysis showed that the protein level of STAT3 was positively correlated with FAP and COL1A1 in our hospital's ovarian cancer tissue chips (r=0.47, P<0.001; r=0.30, P=0.006), the analysis of GEO database GSE26712 dataset showed that the expression of STAT3 gene and FAP, COL1A1 gene were also significantly positively correlated (r=0.31, P<0.001; r=0.52, P<0.001). Conclusion: STAT3 and CAF could promote chemotherapy resistance of ovarian cancer and lead to poor prognosis.

[Effects of visual restoration on dynamic plantar pressure features in elder individuals].

OBJECTIVE: To analyze the effects of visual restoration after cataract surgery on plantar pressure and biomechanics of foot in elder individuals. METHODS: Thirty-two patients [male/female 5/27, (70.1±5.2) years old] with age-related cataract were recruited between October 2016 and December 2019. The footscan system was employed to record the data of plantar pressure during level walking before and 1-month after the cataract surgery. Parameters of peak pressure (PP), impulse (I), pressure-time integral (PTI) and time to peak pressure (TPP) from the regions of the 1st toe (T1), 2nd to 5th toes (T2-5), 1st to 5th metatarsal heads (M1-M5), midfoot (MF), medial hindfoot (HM) and lateral hindfoot (HL) were analyzed respectively. RESULTS: Post-operatively, the visual function was effectively reconstructed with improved visual acuity in both eyes (Z=-4.878, -4.801; P < 0.001). The PP (t=2.266, P=0.031) and I (t=2.152, P=0.039) values in M2 region on the dominant side (right foot) increased statistically at post-operative phase, while the changes of pressure and temporal para-meters in other regions remained stable. There was laterality in plantar pressure at pre-operative phase, manifested as greater PP values in M1, M2, MF, and HM regions on the dominant sides (t=-2.414, -2.478, -2.144, -5.269; P < 0.05), greater PP values in T1, M3, M5 and HL regions on the non-dominant sides (t=4.830, 3.155, 2.686, 3.683; P < 0.05), greater I values in M1, MF, and HM regions on the dominant sides (t=-2.380, -2.185, -5.320; P < 0.05) and greater I values in T1, M3, M5 and HL regions on the non-dominant sides (t=4.489, 2.247, 2.838, 3.992; P < 0.05). post-operatively, the pressure tended to be compatible between the two sides in regions of M3 and MF, while the magnitude of laterality in regions of M1 (ZPP△= -2.721, P=0.007; ZI△=-2.581, P=0.010), M2 (ZPP△=-2.674, P=0.007; ZI△=-2.375, P=0.018) and M5 (ZPP△=1.991, P=0.046; ZI△=2.150, P=0.032) was further increased. CONCLUSION: Changes in plantar pressure after cataract surgery were characterized as increased pressure in the 2nd metatarsal head area on the dominant side. Visual restoration might intensify the laterality in the medial of forefoot on the dominant side and the lateral of forefoot on the non-dominant side.

Reduced Orthodontic Tooth Movement in Enpp1 Mutant Mice with Hypercementosis.

Previous studies revealed that cementum formation is tightly regulated by inorganic pyrophosphate (PPi), a mineralization inhibitor. Local PPi concentrations are determined by regulators, including ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), which increases PPi concentrations by adenosine triphosphate hydrolysis. Orthodontic forces stimulate alveolar bone remodelling, leading to orthodontic tooth movement (OTM). To better understand how disturbed mineral metabolism and the resulting altered periodontal structures affect OTM, we employed Enpp1 mutant mice that feature reduced PPi and increased cervical cementum in a model of OTM induced by a stretched closed-coil spring ligated between the maxillary left first molar and maxillary incisors. We analyzed tooth movement, osteoclast/odontoclast response, and tooth root resorption by micro-computed tomography, histology, histomorphometry, and immunohistochemistry. Preoperatively, we noted an altered periodontium in Enpp1 mutant mice, with significantly increased periodontal ligament (PDL) volume and thickness, as well as increased PDL-bone/tooth root surface area, compared to wild-type (WT) controls. After 11 d of orthodontic treatment, Enpp1 mutant mice displayed 38% reduced tooth movement versus WT mice. Molar roots in Enpp1 mutant mice exhibited less change in PDL width in compression and tension zones compared to WT mice. Root resorption was noted in both groups with no difference in average depths, but resorption lacunae in Enpp1 mutant mice were almost entirely limited to cementum, with 150% increased cementum resorption and 92% decreased dentin resorption. Osteoclast/odontoclast cells were reduced by 64% in Enpp1 mutant mice, with a predominance of tartrate-resistant acid phosphatase (TRAP)-positive cells on root surfaces, compared to WT mice. Increased numbers of TRAP-positive cells on root surfaces were associated with robust immunolocalization of osteopontin (OPN) and receptor-activator of NF-κB ligand (RANKL). Collectively, reduced response to orthodontic forces, decreased tooth movement, and altered osteoclast/odontoclast distribution suggests Enpp1 loss of function has direct effects on clastic function/recruitment and/or indirect effects on periodontal remodeling via altered periodontal structure or tissue mineralization.

Clinical outcome of Descemet stripping automated endothelial keratoplasty in 18 cases with iridocorneal endothelial syndrome.

PurposeTo evaluate the clinical outcome of Descemet stripping automated endothelial keratoplasty (DSAEK) in eyes with iridocorneal endothelial (ICE) syndrome.Patients and methodsA retrospective case series study was conducted. Eighteen consecutive Chinese patients with 20 DSAEK grafts were enrolled. Participants were evaluated by anterior segment optical coherence tomography and confocal microscopy. Postoperative complications, graft survival, endothelial cell counts, corneal thickness, and anterior chamber depth were analysed. A Log-rank test in a Kaplan-Meier analysis and a Cox proportional hazard regression were used to analyse potential risk factors of graft failure.ResultsThe mean follow-up duration was 19.0±8.6 months. The donors' endothelial cell density (ECD) (cells/mm2) values at 1, 3, 6, 12, 18, and 24 months were 3342.2±287.0, 1897.6±745.4, 1793.6±755.7, 1618.1±604.3, 1421.9±650.8, 1265.1±844.1, and 1148.2±1217.8, respectively. Eleven of the 20 grafts exhibited secondary graft failure, with a mean estimated graft survival of 23.4 months. Immediate postoperative complications (air bubble ventilation for elevated intraocular pressure or rebubbling for graft detachment) were more common in eyes exhibiting graft failure (P=0.040). Postkeratoplasty glaucoma surgery emerged as a risk factor of graft failure, with a hazard ratio of 5.174. Eyes with a poor prognosis showed statistically greater central corneal thickness at 1 month, greater graft thickness at 3 months, and a shallower anterior chamber at 6 and 12 months.ConclusionsThe long-term outcome of DSAEK in eyes with ICE syndrome is relatively poor. Immediate postoperative complications, postkeratoplasty glaucoma surgery, thicker corneal parameters, and a shallow anterior chamber were all associated with graft failure.

[Outcomes of total deafness type of idiopathic suddendeafness in different ages].

Objective:This retrospective study was aimed to investigate the characteristics of hearing recovery in the complete deafness type of SSNHL(≥81 dBHL) in patients with different ages.Method:Clinical outcomes of 179 total deafness type of idiopathic sudden deafness were compared.Patients were divided into 5 groups according to age,they were,pediatric group(13 years or less),youthful group(14-44 years),middle-aged group(45-59 years),presenium group(60-74 years),senectitude group(75 years or higher).Patients were divided into 3 groups according to the initial degree of hearing loss: 81 dB group (81-89 dBHL),90 dB group(90- 99 dBHL),100 dB group(100 dBHL or higher).Routine comprehensive treatment including corticosteroids,the inner ear microcirculation improvement drugs,neurotrophic drugs,saturationoxygen and hyperbaric oxygen therapy,etc.was applied.Patients were treated in accordance with the age and body weight.Result:The percentage of youthful group(83/179,46.4%) was highest(P<0.05),middle-aged group(57/179,31.8%)followed(P<0.05),presenium group(26/179,14.5%)was lower(P<0.05),pediatric group(8/179,4.5%) and senectitude group(5/179,2.8%)were the lowest.No a complete recovery in either pediatric group or senectitude group.A complete recovery was rare in the other groups.Recovery rate of the different aged groups was similar(P>0.05).The percentage of 100 dB group(108/179,60.3%) was highest(P<0.05).The percentage of 81 dB group(39/179,21.8%)was similar to 90 dB group(32/179,17.9%)(P>0.05).Recovery rate was similar in 81 dB group(25/39,64.1%)and 90 dB group(18/32,56.2%)(P>0.05).Recovery rate of both 81 dB group and 90 dB group were greater than 100 dB group(24/108,22.2%)(P<0.05).The 100 dB group reduced the satisfactory recovery effects.There were no differences in the proportion of the patients with dizziness(95/179,53.1%)and without dizziness(84/179,46.9%)(P>0.05).Recovery rate of patients without dizziness(43/84,51.2%) was greater than with dizziness(24/95,25.3%)(P<0.05).The percentage of the patients without dizziness(31/39,79.5%)in 81 dB group was the highest(P<0.05),90 dB group(18/32,56.2%)followed(P<0.05).The percentage of the patients with dizziness in 100 dB group(73/108,67.6%)was highest(P<0.05).Recovery rate was similar in the patients without dizziness of 81 dB group(21/31,67.7%)and 90 dB group(11/18,61.1%)(P>0.05).Recovery rate of the above two groups was greater than that of 100 dB group(11/35,31.4%)(P<0.05).Conclusion:Recovery rate of the different aged groups was similar.The percentage of the patients with dizziness in 100 dB group was highest.Initial hearing threshold in excess of 100 dB reduced the satisfactory recovery in patients with total deafness type of SSNHL.Our results provided a good reference for other clinicians.

Mineralization defects in cementum and craniofacial bone from loss of bone sialoprotein.

Bone sialoprotein (BSP) is a multifunctional extracellular matrix protein found in mineralized tissues, including bone, cartilage, tooth root cementum (both acellular and cellular types), and dentin. In order to define the role BSP plays in the process of biomineralization of these tissues, we analyzed cementogenesis, dentinogenesis, and osteogenesis (intramembranous and endochondral) in craniofacial bone in Bsp null mice and wild-type (WT) controls over a developmental period (1-60 days post natal; dpn) by histology, immunohistochemistry, undecalcified histochemistry, microcomputed tomography (microCT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and quantitative PCR (qPCR). Regions of intramembranous ossification in the alveolus, mandible, and calvaria presented delayed mineralization and osteoid accumulation, assessed by von Kossa and Goldner's trichrome stains at 1 and 14 dpn. Moreover, Bsp(-/-) mice featured increased cranial suture size at the early time point, 1 dpn. Immunostaining and PCR demonstrated that osteoblast markers, osterix, alkaline phosphatase, and osteopontin were unchanged in Bsp null mandibles compared to WT. Bsp(-/-) mouse molars featured a lack of functional acellular cementum formation by histology, SEM, and TEM, and subsequent loss of Sharpey's collagen fiber insertion into the tooth root structure. Bsp(-/-) mouse alveolar and mandibular bone featured equivalent or fewer osteoclasts at early ages (1 and 14 dpn), however, increased RANKL immunostaining and mRNA, and significantly increased number of osteoclast-like cells (2-5 fold) were found at later ages (26 and 60 dpn), corresponding to periodontal breakdown and severe alveolar bone resorption observed following molar teeth entering occlusion. Dentin formation was unperturbed in Bsp(-/-) mouse molars, with no delay in mineralization, no alteration in dentin dimensions, and no differences in odontoblast markers analyzed. No defects were identified in endochondral ossification in the cranial base, and craniofacial morphology was unaffected in Bsp(-/-) mice. These analyses confirm a critical role for BSP in processes of cementogenesis and intramembranous ossification of craniofacial bone, whereas endochondral ossification in the cranial base was minimally affected and dentinogenesis was normal in Bsp(-/-) molar teeth. Dissimilar effects of loss of BSP on mineralization of dental and craniofacial tissues suggest local differences in the role of BSP and/or yet to be defined interactions with site-specific factors.

The treatment of port wine stains with a dye laser: a study of 644 patients.

Between June 1989 and December 1996, the flashlamp pumped pulsed dye laser was used to treat port wine stains (PWS) in 644 patients, age range 3 months-93 years (mean 21). The efficacy of the treatment was assessed after more than one year of follow-up. Each factor that might affect the efficacy was then evaluated statistically. Broad lesions required more laser treatments than narrow lesions and clearing tended to start from the periphery of the lesion, indicating the three-dimensional depth of the broader PWS. Patients who had been given previous treatments such as argon laser required about two more laser treatments than those who had not, but there were no clear differences in the efficacy of dye laser treatment between the two groups.

De-epithelialised anterior (anterolateral and anteromedial) thigh flaps for dead space filling and contour correction in head and neck reconstruction.

Anterior (anterolateral and anteromedial) thigh flaps based on the descending branch of the lateral circumflex femoral vessels provide a long vascular pedicle and a large flap without sacrificing main vessels and muscles. Twenty-eight de-epithelialized anterior thigh flaps were transferred for reconstruction of head and neck defects following tumour ablation. Two flaps were lost in patients that had previously undergone high-dose radiotherapy following free tissue transfer. Vascularised fibula, vascularised iliac bone and other tissues were combined with anterior thigh flaps in 13 cases utilising the distal end or derivative branches of the vascular pedicle. Salivary fistula was seen in only one case, although there were many minor and major complications. In five cases, double skin flaps were harvested from the ipsilateral thigh. One of these flaps was used for coverage of intraoral defects, while the other was placed in the submandibular area to fill dead space. Compared with other methods, this multi-flap method is considered to be most suitable for dead space filling and contour correction.

Combined anterior thigh flaps and vascularised fibular graft for reconstruction of massive composite oromandibular defects.

Six massive, composite oromandibular defects were reconstructed using combined anterior (anterolateral and anteromedial) thigh flaps and vascularised fibular graft in bridge or chimeric fashion. Except for minor dehiscence in one case and infection in another, all flaps survived without complication. Anterior thigh flaps provide large-calibre, long vascular pedicles while derivative branches from the lateral circumflex femoral system facilitate simultaneous transplantation of multiple components. Because the pedicle of the vascularised fibular graft is of insufficient length, the lateral circumflex femoral vessels provide a remote vascular source as a flow-through vascular conduit. A combined flap using the lateral circumflex femoral system is considered to be most suitable for reconstruction of through-and-through defects of the head and neck. The authors describe the advantages of this method and the detailed anatomy of the cutaneous perforators of the anterior thigh flaps.

Perforator-based flap for coverage of lumbosacral defects.

Lumbosacral defects on 20 patients were covered with a perforator-based flap. Cutaneous perforators derived from the 9th and 10th intercostal arteries, the 4th lumbar artery, and multiple gluteal perforators that penetrate the gluteus maximus muscle were used as vascular pedicles. Minor complications occurred in five cases. Using this method, minimal morbidity of the donor site is expected because the gluteus maximus need not be sacrificed. Accordingly, perforator-based flaps are especially indicated for ambulatory patients, but for paraplegic patients as well. Even in the event of recurrence, another perforator-based or musculocutaneous flap can be elevated from the ipsilateral side because of the presence of multiple perforators in the lumbosacral and gluteal regions.

Reconstruction of posttraumatic defects of the foot by flow-through anterolateral or anteromedial thigh flaps with preservation of posterior tibial vessels.

Massive posttraumatic defects of the foot in 4 patients and a tibial malunion in another were repaired by flow-through anterior (anterolateral and anteromedial) thigh flaps. Posterior tibial vessels were used as the recipient vessel just below the level of the medial malleolus. Soft-tissue defects in foot-injured patients were covered with flow-through anterior thigh flaps interposing the descending branch of the lateral circumflex femoral system between the transected posterior tibial vessels. In another patient, a combined anteromedial thigh flap and vascularized iliac bone cross-leg "chimeric" flap were successfully transferred for the reconstruction of a malunion of a tibial fracture, which had been accompanied by severe vascular damage to the thigh. The utilization of derivative branches from the lateral circumflex femoral system facilitates revascularization of ischemic areas and simultaneous transplantation of multiple components, as well as preserves recipient vessels. Owing to the stable blood supply, these flaps can be readily processed into both deepithelialized and thin flaps.