BACKGROUND:Chondrocyte apoptosis is an important factor in the development of osteoarthritis,and Complanatoside A has a flavonoid effect,which can inhibit apoptosis of various cells,but its effect on chondrocyte apoptosis and the mechanism of action are not clear. OBJECTIVE:To investigate the intrinsic association and mechanism of Complanatoside A in chondrocyte apoptosis based on the Wnt/β-catenin signaling pathway. METHODS:(1)The cartilage tissues of the femur and tibia transected during knee arthroplasty were collected,and chondrocytes were isolated,cultured in vitro,and identified.(2)Cell counting kit-8 was used to detect the optimal intervention concentration of Complanatoside A in the concentration range of 0-160 μmol/L.(3)Chondrocytes were divided into blank group,sodium nitroprusside(1.5 mmol/L)-induced group,and sodium nitroprusside(1.5 mmol/L)+Complanatoside A(5 μmol/L)group.The viability and apoptosis rate of the cells in each group were detected by cell counting kit-8 and flow cytometry.The expression of type Ⅱ collagen and SOX9 was detected by immunofluorescence staining.The expression of apoptosis-related proteins and Wnt/β-catenin pathway proteins was detected by western blot assay. RESULTS AND CONCLUSION:The cells extracted in vitro were cultured and stained,and were clearly identified as chondrocytes.Complanatoside A had no obvious cytotoxicity to chondrocytes in the concentration range of 0-80 μmol/L,and significantly improved the chondrocyte viability in the concentration range of 2.5-10 μmol/L,especially when the concentration was 5 μmol/L.The apoptotic rate of chondrocytes was higher in the sodium nitroprusside-induced group than the blank control group,while the apoptotic rate was lower in the sodium nitroprusside+Complanatoside A group than the sodium nitroprusside-induced group.The fluorescence intensity of type Ⅱ collagen and SOX9 in chondrocytes was weaker in the sodium nitroprusside-induced group than the blank control group,while the fluorescence intensity of type Ⅱ collagen and SOX9 in the sodium nitroprusside+Complanatoside A group was higher than that of the sodium nitroprusside-induced group.In the sodium nitroprusside-induced group,the protein expression of Bax,Caspase-3,matrix metalloproteinase 13,Wnt3a,Wnt5a and β-catenin was higher than that of the blank control group,while the protein expression of Bcl-2 was lower than that of the blank control group.In the sodium nitroprusside+Complanatoside A group,except for the protein expression of Bcl-2 which was higher than that of the sodium nitroprusside-induced group,the expression of the other aforementioned proteins was lower than that of the sodium nitroprusside-induced group.To conclude,Complanatoside A has a certain inhibitory effect on chondrocyte apoptosis,which could regulate apoptosis-related proteins and promote the expression of chondrocyte regulatory factors,and presumably might play a role through inhibiting the Wnt/β-catenin signaling pathway.

OBJECTIVE To study the inhibitory effect mechanism of rhynchophylline solid lipid nanoparticles （Rhy-SLN） on the proliferation of airway smooth muscle cells （ASMCs） in asthmatic model mice. METHODS Asthma model was prepared by ovalbumin+calmogastrin sensitization. The primary isolation and culture of ASMCs were performed， and morphological observation and identification were also conducted [when α -smooth muscle actin （α -SMA） appeared red and Desmin appeared green in ASMCs， indicating successful cultivation of ASMCs]. The cells were divided into blank group （ASMCs of normal mice）， model group （ASMCs of asthma model mice）， Rhy-SLN group （ASMCs of asthma model mice）， recombinant suppressors of cytokine signaling 1 （SOCS1） overexpression group （ASMCs of asthma model mice transfected with SOCS1 vector）， SOCS1-RNAi group （ASMCs of asthma model mice transfected with SOCS1-RNAi vector） and SB203580 group [p38 mitogen-activated protein kinase （p38 MAPK） inhibitor， ASMCs of asthma model mice]. The cells of each group were added into the corresponding culture medium containing drug （10 μmol/L） or not containing drug for 24 hours. MTT method was used to detect the proliferation of ASMCs in asthmatic mice； Western blot assay was used to detect the protein expressions of α-SMA， interleukin-1β （IL-1β）， SOCS1， p38 MAPK and phosphorylated p38 MAPK （p-p38 MAPK） in ASMCs. RESULTS The primary ASMCs of mice varied in shape and size， presenting irregular， spindle and triangular shapes；α-SMA appeared red and Desmin appeared green， indicating successful cultivation of ASMCs. Compared with model group， ASMCs absorbance values and protein expressions of α -SMA， p38 MAPK， and p-p38 MAPK were reduced significantly in Rhy- SLN group， SOCS1 overexpression group and SB203580 E-mail：wangmeng106@163.com group， while protein expression of SOCS1 （except for group） was increased significantly （P＜0.05）； protein expressions of IL-1β was reduced significantly in ASMCs （P＜ 0.05）. ASMCs absorbance values and protein expressions of α-SMA， SOCS1， p38 MAPK and p-p38 MAPK were increased significantly in SOCS1-RNAi group （P＜0.05）. CONCLUSIONS Rhy-SLN can inhibit the proliferation of ASMCs， the mechanism of which may be associated with overexpression of SOCS1 and inhibiting the protein expressions of IL-1β and p38 MAPK.

Neurogenesis decline in hippocampal dentate gyrus (DG) participates in stress-induced depressive-like behaviors, but the underlying mechanism remains poorly understood. Here, we observed low-expression of NOD-like receptor family pyrin domain containing 6 (NLRP6) in hippocampus of stress-stimulated mice, being consistent with high corticosterone level. NLRP6 was found to be abundantly expressed in neural stem cells (NSCs) of DG. Both Nlrp6 knockout (Nlrp6^-/-) and NSC-conditional Nlrp6 knockout (Nlrp6CKO) mice were susceptible to stress, being more likely to develop depressive-like behaviors. Interestingly, NLRP6 was required for NSC proliferation in sustaining hippocampal neurogenesis and reinforcing stress resilience during growing up. Nlrp6 deficiency promoted esophageal cancer-related gene 4 (ECRG4) expression and caused mitochondrial dysfunction. Corticosterone as a stress factor significantly down-regulated NLRP6 expression, damaged mitochondrial function and suppressed cell proliferation in NSCs, which were blocked by Nlrp6 overexpression. ECRG4 knockdown reversed corticosterone-induced NSC mitochondrial function and cell proliferation disorders. Pioglitazone, a well-known clinical drug, up-regulated NLRP6 expression to inhibit ECRG4 expression in its protection against corticosterone-induced NSC mitochondrial dysfunction and proliferation restriction. In conclusion, this study demonstrates that NLRP6 is essential to maintain mitochondrial homeostasis and proliferation in NSCs, and identifies NLRP6 as a promising therapeutic target for hippocampal neurogenesis decline linked to depression.

Objective:To propose automatic measurement of global and local tessellation density on color fundus images based on a deep convolutional neural network (DCNN) method.Methods:An applied study. An artificial intelligence (AI) database was constructed, which contained 1 005 color fundus images captured from 1 024 eyes of 514 myopic patients in the Northern Hospital of Qingdao Eye Hospital from May to July, 2021. The images were preprocessed by using RGB color channel re-calibration method (CCR algorithm), CLAHE algorithm based on Lab color space, Retinex algorithm for multiple iterative illumination estimation, and multi-scale Retinex algorithm. The effects on the segmentation of tessellation by adopting the abovemetioned image enhancement methods and utilizing the Dice, Edge Overlap Rate and clDice loss were compared and observed. The tessellation segmentation model for extracting the tessellated region in the full fundus image as well as the tissue detection model for locating the optic disc and macular fovea were built up. Then, the fundus tessellation density (FTD), macular tessellation density (MTD) and peripapillary tessellation density (PTD) were calculated automatically.Results:When applying CCR algorithm for image preprocessing and the training losses combination strategy, the Dice coefficient, accuracy, sensitivity, specificity and Jordan index for fundus tessellation segmentation were 0.723 4, 94.25%, 74.03%, 96.00% and 70.03%, respectively. Compared with the manual annotations, the mean absolute errors and root mean square errors of FTD, MTD, PTD automatically measured by the model were 0.014 3, 0.020 7, 0.026 7 and 0.017 8, 0.032 3, 0.036 5, respectively.Conclusion:The DCNN-based segmentation and detection method can automatically measure the tessellation density in the global and local regions of the fundus of myopia patients, which can more accurately assist clinical monitoring and evaluation of the impact of fundus tessellation changes on the development of myopia.

Abstract Oral health problems is one of major public health problems that threatens children s overall health. Children s oral health is closely related to family parenting styles. Currently, grandparenting is becoming increasingly common in China, and its impact on children s oral health should not be underestimated. This article reviews the impacts of the oral health knowledge level, behavior habits, gender, socioeconomic status, and other factors related to grandparenting on children s oral health, in order to reduce the negative impact of grandparenting involvement on children s oral health, and to provide theoretical basis and guidance.

Diabetic patients with poor glycemic control are exposed to media containing mold spores, and spores enter the body, which may lead to refractory infections. This article combines case and literature reviews, proposes the diagnosis and treatment method of mold infection, and provides some guidances for subjects who long-term exposure to mold groups such as farmers and immunocompromised people.

Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.

Objective To evaluate the efficacy and safety of recombinant human endostatin (rh-endostatin) combined with cis-platinum for patients with malignant pleural effusions.Methods A computer-based online search was performed through Elsevier, PubMed, Medline, Cochrane Library, China National Knowledge Infrastructure (CNKI), Chinese BioMedical Literature Database and Wanfang Database.According to the inclusion and exclusion criteria, the randomized controlled trials about short-term therapeutic effect of rh-endostatin combined with cis-platinum on malignant pleural effusions published until December 2015 were selected.Quality of the studies was assessed using modified Jadad scale.After data extraction, a Meta-analysis was performed by RevMan 5.3 software.Relative risk (RR) and its 95% confidence interval (CI) were calculated.The Egger test was performed by Stata 12.0 software.Results Fourteen eligible randomized controlled trials were included in this Meta-analysis involving 1 330 patients,665 in cis-platinum alone group(control group), and 665 in rh-endostatin combined with cis-platinum group(treatment group).The results showed that there were significant improvements in overall response rate (ORR) [73.53% vs 45.41%,RR=1.62,95%CI(1.47,1.78),P<0.000 01] and the rate of quality of life improvement [71.65% vs 46.94%,RR=1.52,95%CI(1.38,1.68),P<0.000 01] in the treatment group, as compared with those of the control group.Meanwhile, there were no statistically significant differences in the rate of cardio toxicity [10.38% vs 5.77%,RR=1.73,95%CI(0.99,3.03),P=0.06].Conclusion This Meta-analysis indicated that in comparison with cis-platinum alone, rh-endostatin combined with cis-platinum has a better therapeutic effect on malignant pleural effusions.

Objective To explore the effects of improvements on setup error ( SE ) and clinical target ( CTV ) margin of supraclavicular field ( SCF ) by using moisture-cured resin cushion and breast bracket for lower neck fixation in breast cancer patients who underwent post-mastectomy radiotherapy. Methods Totally 13 patients with breast cancer who underwent post-mastectomy radiotherapy were enrolled. All patients were immobilized by breast bracket and moisture-cured resin cushion. Firstly, each patient′s lower neck and head was fixed well by moisture-cured resin cushion, filling the gap between the neck and breast bracket. Secondly,each patient underwent the cone-beam CT ( CBCT) at the first, tenth and twentieth treatment after positioning. Then these CBCT images were registered to the planning CT to determine setup errors in translational and rotational directions, and SCF CTV margins by the systematic and random errors were evaluated. Results The setup errors in x (left-right), y (superior-inferior), z (anterior-posterior) translational directions were (2. 16 ± 1. 25), (1. 50 ± 1. 28), (1. 94 ± 1. 12) mm and (1. 76 ±1. 87)°, (1. 82 ±1. 12)°, (0. 99 ±0. 58)°, respectively in θ (pitch degree),Ф(roll degree),ψ( yaw degree) rotational directions. Non-parametric rank test ( Mann-Whitney U test) was performed with previous data, the differences of the setup error in y, z,θ directions were statistically significant ( Z =4. 152, 3. 415, 2. 053, P<0. 05). The margins from CTV were 4. 07, 4. 03 and 3. 73 mm in x, y and z directions, respectively. Compared with the previous study on SCF, CTV margin required 8, 8 and 6 mm in x, y and z axis directions, the volume of CTV to PTV were decreased by 32. 73% on average. Conclusions Compared with plastic circular pillow alone, moisture-cured resin cushion with breast bracket locating method could reduce setup errors in SCF target region. The margins from CTV to PTV weredecreased to 4. 07, 4. 03 and 3. 73 mm in x, y and z axis directions at least.

OBJECTIVETo evaluate the role and value of Changhai fulcrum bending radiograph(CH-FBR) in curve flexibility assessment of adolescent idiopathic scoliosis(AIS) patients.

METHODSThirty-seven AIS patients treated between June 2012 and August 2013 were enrolled, including 31 female and 6 male patients whose age ranged from 10 to 19 years, averaged of 15.0 years. The assessment of radiographs included preoperative standing posterior-anterior radiograph, supine side-bending radiograph, traditional fulcrum bending radiograph, Changhai fulcrum bending radiograph and postoperative standing posterior-anterior radiograph. Postoperatively, radiographs were assessed at one week. The CH-FBR was performed at the lowest height and the optimized height which means the weight on the fulcrum touch the maximum. All measurements of angle were made with use of the Cobb method. The flexibility of the curve as well as the correction rate and fulcrum bending correction index (FBCI) were calculated for all patients. The maximum height of CH-FBR, basic weight and maximum weight were measured for all AIS. Paired t-tests were used to assess differences between preoperative and postoperative curves within group samples. The Pearson correlation coefficients were calculated using bivariate analysis between CH-FBR flexibility rate and correction rate, the maximum height of CH-FBR and maximum weight, the height changes of CH-FBR and weight changes.

RESULTSA total of 46 curves were involved in this study, including 28 thoracic and 18 thoracolumbar/lumbar curves. Preoperatively, the mean Cobb angle of the 46 structural curves was 47° ± 11°. Postoperatively, the mean Cobb angle was 11° ± 5°. Cobb's angle in supine side-bending(t = 7.2, P = 0.001), traditional fulcrum bending (t = 7.1, P = 0.001) and lowest height of Changhai fulcrum bending (t = 6.5, P = 0.001) were significantly different from the postoperative Cobb angle; Cobb's angle in traditional FBR (t = 11.0, P = 0.001) and lowest height of Changhai fulcrum bending (t = 13.6, P = 0.001) were significantly different from the optimized height CH-FBR Cobb angle. There was no significant difference found between traditional FBR Cobb angle and lowest height CH-FBR Cobb angle (t = 2.0, P = 0.051), optimized height CH-FBR Cobb angle and postoperative Cobb angle (t = 0.9, P = 0.36), lowest height CH-FBR Cobb angle and traditional FBR Cobb angle(t = 2.0, P = 0.051). The maximum height of CH-FBR, basic weight and maximum weight were (29.6 ± 1.4)cm,(20 ± 6)kg, and (40 ± 6) kg. Preoperatively, the mean Cobb angle of the 28 structural curves(main thoracic curves) was 46° ± 11°. Postoperatively, the mean Cobb angle was 12° ± 6°. Preoperatively, the mean Cobb angle of the 18 structural curves(thoracolumbar/lumbar curves) was 49° ± 12°. Postoperatively, the mean Cobb angle was 10° ± 5°. The results were same in 28 structural curves, 18 structural curves as well as 46 curves. Correlation analysis of 46 curves indicated that the maximum height of CH-FBR positively correlated with maximum weight (r = 0.69, r(2) = 0.47, P = 0.001), the height changes of CH-FBR positively correlated with weight changes on CH-FBR (r = 0.62, r(2) = 0.38, P = 0.001).

CONCLUSIONSCH-FBR is a more reliable and effective method than traditional FBR and supine side-bending for curve flexibility evaluation in AIS patients. Moreover, compared to the traditional FBR and side-bending radiograph, the flexibility suggested by the optimized height CH-FBR more closely approximates the postoperative result made by pedicle screws fixation and fusion.

Adolescent ; Child ; Female ; Humans ; Male ; Prospective Studies ; Radiography ; Range of Motion, Articular ; Scoliosis ; diagnostic imaging ; surgery ; Young Adult