Acute generation of reactive oxygen species that induced by doxycycline pretreatment results in rapid cell death in polyphyllin G-treated osteosarcoma cell lines.

Polyphyllin G, a pennogenyl saponin extracted from Paris polyphylla, has been shown to possess antitumor effects. In this study, we demonstrated that doxycycline, an antibiotic medicine, could significantly enhance the sensitivities of osteosarcoma cell lines to polyphyllin G. As the cells were pretreated with doxycycline at non-toxic concentrations and then co-exposed to polyphyllin G, this combination could induce a rapid cell death distinct from apoptosis. The non-apoptotic cell death was characterized by a loss of integrity of plasma membrane without externalization of phosphatidyl serine. Furthermore, this combined treatment resulted in suppression of cell viability and colony-forming ability, and increased the level of γ-H2A.X, a critical marker for DNA damage, in osteosarcoma cell lines. When examining the underlying mechanism, it was revealed combination of polyphyllin G and doxycycline triggered an enhanced generation of reactive oxygen species (ROS), and up-regulated mitochondrial oxidative stress within 0.5 h. Co-administration of the ROS inhibitor NAC reversed the suppressed cell viability and colony-forming ability, and abolished the increased level of γ-H2A.X in the cells with the combined treatment, indicating that the enhanced ROS was involved in the anti-proliferative effect of the combined treatment. Overall, the results demonstrated that doxycycline may function as chemosensitizers by inducing an acute and lethal ROS production to enhance cytotoxic of polyphyllin G in osteosarcoma cell lines, and the combined use of drugs may provide an alternative thinking for the development of new therapeutic agents.

Clinical feature and predictive factor analysis for spontaneous regression of retinopathy of prematurity in a Chinese population.

AIM: To investigate the ratio of spontaneous regression of retinopathy of prematurity (ROP) and to explore the possible relevant predictive factors. METHODS: A retrospective review of 405 infants who were diagnosed with ROP and mother during pregnancy were collected. Stage, zone, and duration of ROP were recorded. Statistical analysis was performed on 51 possible predictive factors. RESULTS: Totally 356 infants showed spontaneous regression. The incidence was 100%, 95.3%, and 22.7% in stage 1, 2, and 3, respectively. The 13.4% of the ROP with plus disease eventually resolved spontaneously. All affected eyes of aggressive posterior retinopathy of prematurity (AP-ROP) failed to spontaneously regress. The mean duration of ROP was 7.2wk in patients with spontaneous resolution of ROP. Days of mechanical ventilation (OR=0.981, 95%CI, 0.965-0.997, P=0.021), retinal hemorrhage (OR=0.173, 95%CI, 0.064-0.470, P=0.001), delivery pattern (OR=2.750, 95%CI, 1.132-6.681, P=0.025), maternal anemia in pregnancy (OR=0.142, 95%CI, 0.036-0.563, P=0.005), the stages (at initial diagnosis OR=0.183, 95%CI, 0.041-0.816, P=0.026; at final diagnosis OR=0.031, 95%CI, 0.006-0.167, P<0.001), and with plus disease or not (OR=0.005, 95%CI, 0.001-0.031, P<0.001) were independent predictive factors of the spontaneous regression of ROP. CONCLUSION: Most mild ROP can spontaneously resolve. Active treatment is still recommended for stage 3 ROP, zone I ROP, AP-ROP, and ROP with plus disease. Prolonged mechanical ventilation and concurrent retinal hemorrhage reduce the likelihood of spontaneous ROP resolution. The pattern of delivery and the mother's anemia during pregnancy can also affect the prognosis of ROP.

A One-Step, Monolayer Culture and Chemical-Based Approach to Generate Insulin-Producing Cells From Human Adipose-Derived Stem Cells to Mitigate Hyperglycemia in STZ-Induced Diabetic Rats.

The global population of individuals afflicted with diabetes mellitus has been increasing year by year, and this disease poses a serious threat to human health as well as the economies worldwide. Pancreatic or islet transplantations provide one of the most effective and long-term therapies available to treat diabetes, but the scarcity and quality of pancreatic islets limit their use in treatments. Here, we report the development of a one-step, monolayer culture, and chemical-based protocol that efficiently mediates the differentiation of human adipose-derived stem cells (hADSCs) into insulin-producing cells (IPCs). Our data indicate that hADSCs in monolayer culture that are allowed to differentiate into IPCs are superior to those in suspension cultures with respect to insulin secretion capacity (213-fold increase), cell viability (93.5 ± 3.27% vs. 41.67 ± 13.17%), and response to glucose stimulation. Moreover, the expression of genes associated with pancreatic lineage specification, such as PDX1, ISL1, and INS (encoding insulin), were expressed at significantly higher levels during our differentiation protocol (6-fold for PDX1 and ISL1, 11.5-fold for INS). Importantly, in vivo studies demonstrated that transplantation with IPCs significantly mitigated hyperglycemia in streptozotocin-induced diabetic rats. Our results indicate that this one-step, rapid protocol increases the efficiency of IPC generation and that the chemical-based approach for IPC induction may reduce safety concerns associated with the use of IPCs for clinical applications, thereby providing a safe and effective cell-based treatment for diabetes.

Engineered cell-laden thermosensitive poly(N-isopropylacrylamide)-immobilized gelatin microspheres as 3D cell carriers for regenerative medicine.

Several studies have focused on using cell carriers to solve the problem of mesenchymal stem cell expansion on regenerative medicine. However, the disadvantages of using prolonged enzymatic treatment and low cell harvest efficiency still trouble researchers. In this study, PNIPAAm-immobilized gelatin microspheres (abbreviated as GNMS) were synthesized using a simple power-driven flow-focusing microinjection system. The developed thermosensitive GNMS can allow easier harvesting of cells from the microspheres, requiring only 10 â€‹min of low-temperature treatment and 5 â€‹min of trypsin treatment. The developed GNMS was characterized by Fourier-transform infrared spectroscopy, optical microscopy, and scanning electron microscopy. Further, live/dead staining, F-actin staining, and PrestoBlue cell viability assays were used to evaluate cytotoxicity, cell morphology, cell proliferation, and harvest efficiency. The gene expression of stem cell markers was determined by real-time quantitative PCR (Q-PCR) analysis to investigate the stemness and phenotypic changes in Wharton's jelly-derived mesenchymal stem cells. The results showed that the engineered cell-laden thermosensitive GNMS could significantly increase the cell harvest rate with over 99% cell survival rate and no change in the cell phenotype. Thus, the described strategy GNMS could be the suitable 3D cell carriers in the therapeutic application and opens new avenues for regenerative medicine.

Early detection of ocular abnormalities in a Chinese multicentre neonatal eye screening programme-1-year result.

PURPOSE: Early detection of ocular abnormalities in newborns is essential for timely diagnosis and treatment. This study aimed to assess the 1-year result of a multicentre prospective neonatal eye examination programme with wide-field digital imaging system in China. METHODS: A multicentre collaborative prospective study group for neonatal eye screening was established in nine hospitals, including eight Maternal and Children's Hospitals, and one general hospital across China from July 2016 to June 2017. Ocular examinations were performed on newborns within 28 days after birth using a wide-field digital imaging system. Data were reviewed and analysed. The primary outcome was the prevalence of ocular abnormalities in neonates. RESULTS: We detected 13 514 (20.91%) abnormal cases in 64 632 newborns. The most frequent abnormality was retinal haemorrhage (RH; 11.83%). Most of mild RH resolved spontaneously. Among those who were beyond retinopathy of prematurity (ROP) screening criteria of China (gestational age ≥32 w and birthweight ≥2000 g), the total number of neonates with ocular abnormality was 12 218/62 799(19.45%). 59.44% of neonatal ocular abnormalities detected (accounting for 11.56% of all the screened population) needed further interference or observation. Among them, 258 patients (0.41% of all the screened population) needed immediate or timely intervention, including congenital cataract, retinal detachment, retinoblastoma and other ocular abnormalities. One thousand and ninety-eight patients (1.75% of all the screened neonates) should be followed up closely and needed further diagnosis or intervention if necessary, such as ROP or ROP-like retinopathy, familial exudative vitreoretinopathy and persistent hyperplasia of primary vitreous. Five thousand nine hundred and six patients (9.4%) with minor clinical significance needed short-term follow-up. CONCLUSIONS: This prospective multicentre study of newborn ocular examination showed a relatively high prevalence of ocular abnormalities. There are a relatively high percentage of congenital eye pathology that required further referral and treatment in those neonates who were not screened routinely. According to the benefits and risks associated with neonatal eye examinations, neonatal ocular screening programme can detect ocular abnormalities at the very early stage and may play a positive role in promoting paediatric eye health.

Asymmetric Outcomes of Type 1 Retinopathy of Prematurity after Bilateral Intravitreal Ranibizumab Treatment.

Purpose. To present cases with retinopathy of prematurity (ROP), who were treated with intravitreal injection of ranibizumab (IVR) and had unpredictable asymmetric outcomes. Methods. A retrospective review was performed in infants with type 1 ROP and had bilateral IVR (0.25 mg/0.025 mL) as initial treatment. Patients were classified into the asymmetric outcome group and the symmetric outcome group. Results. Eighty-four patients (168 eyes) were included. There were 18 eyes of 9 patients (10.7%) in the asymmetric outcome group and 150 eyes of 75 patients (89.3%) in the symmetric outcome group. In the symmetric outcome group, 86 eyes (57.3%) had ROP regression, 60 eyes (40%) had reactivation requiring laser treatment, and 4 eyes (2.7%) progressed to retinal detachment requiring vitrectomy. In the asymmetric outcome group, one of the eyes of the 9 patients had ROP regression with/without reactivation after IVR, while the contralateral eyes had negative response, including remarkable posterior fibrosis, partial or total retinal detachment, and vitreous hemorrhage. There was statistically significant difference between the birth weight of the two groups. Conclusion. Contralateral eyes with ROP can take a different clinical course after ranibizumab treatment. High rate of reactivation after IVR is another concern that ophthalmologists should pay attention to.

Ranibizumab Injection as Primary Treatment in Patients with Retinopathy of Prematurity: Anatomic Outcomes and Influencing Factors.

PURPOSE: To investigate the anatomic outcomes and influencing factors of ranibizumab in the treatment of retinopathy of prematurity (ROP). DESIGN: Retrospective case series. PARTICIPANTS: A total of 283 eyes of 145 patients with type 1 ROP treated with intravitreal injection of ranibizumab (IVR) as primary treatment. METHODS: Retrospective review of infants who were diagnosed with type 1 ROP and accepted IVR (0.25 mg/0.025 ml) as primary treatment from January 2012 to August 2015. The anatomic outcomes and the influencing factors were analyzed. MAIN OUTCOME MEASURES: Anatomic outcomes of ROP eyes after IVR and the influencing factors. RESULTS: A total of 283 eyes of 145 patients were included in this study. There were a total of 266 eyes (94.0%) in the positive response group and 17 eyes (6.0%) in the negative/no response group after IVR. Within the positive response group, 139 eyes (48.6%) were in the regression without reactivation subgroup, and 127 eyes (44.9%) were in the regression with reactivation subgroup. A total of 152 eyes received additional laser or surgical treatment. At the last visit, 278 eyes (98.2%) had attached retinas, and 5 eyes (1.8%) had retinal detachment. A classification tree model showed that for patients with gestational age (GA) ≤29.5 weeks, the possibility of experiencing reactivation after IVR is higher than that of those with GA >29.5 weeks (61.6% vs. 29.6%). Moreover, for patients with GA ≤29.5 weeks, those diagnosed with zone II stage 2+ ROP have a lower possibility of experiencing reactivation than other patients (37.9% vs. 80%). CONCLUSIONS: Intravitreal injection of ranibizumab seemed to be effective in treating patients with ROP. After treatment, there were primarily 3 different outcomes. Our predictive tree model is helpful for ophthalmologists to evaluate the risk of reactivation.

Subsequent vertebral fracture after vertebroplasty: incidence and analysis of risk factors.

STUDY DESIGN: A retrospective review and analysis of consecutive patients who underwent single-level vertebroplasty at our institute between March 2002 and March 2006. OBJECTIVE: To analyze the risk factors for subsequent fractures after vertebroplasty and to predict the postoperative fracture-free time and rate. The effect of bone cement volume injected was also evaluated. SUMMARY OF BACKGROUND DATA: Previous studies of subsequent fractures after vertebroplasty showed conflicting conclusions about risk factors. The frequency of refracture also varied, ranging from 12% to 52%. Most new fractures occurred at adjacent levels, with different risk factors identified. No data were available on the effect of injected bone cement volume, and no consensus had been reached as to the optimal cement volume. METHODS: All enrolled patients were treated with single-level vertebroplasty and followed a standardized postoperative care protocol. Data from medical records and radiographs were collected and analyzed. Variables included patient constitutional factors, radiographic parameters, and volume of injected bone cement. RESULTS: A total of 166 patients (76 men, 90 women) with a mean age of 73.4 years were enrolled in this study. The mean follow-up time was 15.3 months. The overall refracture rate was 38%, with a mean fracture-free interval of 32 months. Both a greater volume of bone cement injected and a greater degree of vertebral height restored contributed significantly to the risk of subsequent adjacent fracture. No risk factor for subsequent remote fracture was identified. A greater volume of bone cement injected was positively correlated with deformity correction after vertebroplasty. CONCLUSION: Most subsequent fractures occurred at the adjacent level within the first 3 months. Patient preoperative condition did not help predict refracture. Although a greater volume of bone cement injected when performing vertebroplasty contributed to the risk of subsequent adjacent fracture, it resulted in a greater improvement of kyphosis.

[Application of near-infrared spectrum technology to research of weathering of red sandstone relics].

In the present paper, with near infrared spectroscopy technology, the weathering mechanism of red sandstone relics was studied. Six groups of red sandstone samples were analyzed using near infrared spectroscopy technology. The results show that the near-infrared spectroscopy technology can analyze the material composition of red sandstone before and after weathering, aiming to explore their components changed. So it is a quick and efficient means of research with characteristic of less measurement sample and speed and non-damage and being pollution-free compared with other research techniques. All the characteristic shows that it is also well for studying other stone cultural relics. Especially for those with sampling difficulty and treasure valuable, non-destruction of stone cultural relics is particularly important. So with time advancing, near infrared technology as a research means of stone relics, its meaning will be more prominent.

Prospective analysis of a new minimally invasive technique for paediatric Gartland type III supracondylar fracture of the humerus.

OBJECTIVE: The purpose of this study was to introduce a minimally invasive surgery using mosquito forceps for achieving the reduction of severely displaced supracondylar fracture of the humerus in children and evaluate the clinical outcome. MATERIALS AND METHODS: Gartland type III supracondylar fractures of the humerus in children under the age of 12 were analysed from January 2000 to January 2007 at our institute. The control group cases were selected by successful reduction using the standard technique described by Rockwood and Wilkins (2001). The others composed the study group. This study included chart reviews, radiographic images of both the antero-posterior and the lateral views, physical examination, and Mayo elbow performance index scores preoperative, postoperative, and final follow-up. RESULTS: Evaluations were completed for 42 cases. The mean age was 6.9 years and the group consisted of 23 boys and 19 girls. The mean follow-up time was 34.5 months. Twenty-six cases were of injuries to the left side, and 16 were of injuries to the right. The Mayo elbow performance index score was excellent in both groups. Operation time in the control and study groups was 46 and 79min, respectively. There was one case of iatrogenic ulnar nerve injury each in the study and control group. One case of loss of reduction and another with a combination of ulnar nerve injury and loss of reduction were noted in the control group. All four cases had an excellent final outcome. No mosquito-forceps-related complication was found, and in the final follow-up, no cubitus varus was found. CONCLUSION: Minimally invasive surgery with the assistance of mosquito forceps for the reduction of severely displaced supracondylar fracture of the humerus in children was shown to be a safe and effective alternative method. The final functional outcome was the same as that of the standard technique.