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.

At present, the major public health challenges caused by novel coronavirus infection have gradually subside. However, a large number of people are still suffering from long-novel coronavirus syndrome or post-novel coronavirus syndrome. The clinical manifestations of long coronavirus syndrome are related to multiple systems, such as respiratory, circulatory, nervous, digestive and musculoskeletal systems, with various long-term persistent symptoms after novel coronavirus infection. At the same time, the infection of the novel coronavirus is an important cause of frailty and sarcopenia in the elderly population. However, at present, the scholars have not paid enough attention to the skeletal muscle weakness caused by the novel coronavirus. Therefore, this paper focuses on the long-novel coronavirus syndrome and sarcopenia to explore the pathological mechanism of skeletal muscle attenuation caused by the SARS-CoV-2 mediated "cytokine storm", mitochondrial damage, hypoxia state and other links, so as to raise the attention of clinical and academic researchers and improve the clinical strategy of frailty and sarcopenia after novel coronavirus infection.

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.

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.

Objective:To investigate the in vitro antibacterial effects of imipenem combined with common antibiotics on bla KPC-2 type carbapenem resistant klebsiella pneumoniae (CRKP) targeting bla KPC-2 gene. Methods:Six strains of unrepeated bla KPC-2 type confirmed by polymerase chain reaction and DNA sequence were isolated in Yueqing People's Hospital, China between January 2018 and January 2019 were included in this study. The susceptibility rate of imipenem against nine conventionally used antibiotics was determined. The sensitivity test of imipenem combined with eight antibiotics was performed with the checkerboard method. Fractional inhibitory concentration was calculated to assess the efficacy of imipenem combined with common antibiotics. The in vitro treatment time-antibacterial effect curve was drawn to evaluate the antibacterial effects. Results:The resistance rate of six strains of bla KPC-2 type was 100.00% (6/6) for imipenem, meropenem, ceftazidime, ciprofloxacin, rifampicin and cefotaxime, and it was 66.67% (4/6) for minocycline and clavulanic acid and 33.33% (2/6) for tigecycline. Imipenem combined with tigecycline had a better antibacterial effect and exhibited a synergistic effect on four strains of bla KPC-2 type CRKP and an additive effect on two strains of Bla KPC-2 type CRKP. The curve of time for in vitro treatment of KPN2 with imipenem combined with tigecycline against bactericidal effect revealed that the antibacterial rate of imipenem at the 1/2 minimum inhibitory concentration combined with tigecycline at the 1/4 minimum inhibitory concentration was > 95% at (t+2) and the antibacterial effect could maintain (t+10) hours to (t+12) hours. The antibacterial rate of imipenem combined with tigecycline against strain 002 was gradually decreased with time, and the growth curve of strain 002 rised gradually. Conclusion:In vitro drug sensitivity test revealed that imipenem combined with tigecycline exhibits a good synergistic effect on bla KPC-2 type CRKP. Findings from this study provide a reference for clinical treatment of bla KPC-2 type CRKP.

Primary ureteral plasmacytoid carcinomas is a rare tumor with high grade and poor diagnosis. Pathological and immunohistochemical staining play an extremely key role in diagnosis since there is no specific clinical and radiological evidence. The surgical removement is the first line treatment. Herein, we report a case of ureteral plasmacytoid carcinoma that was well controlled with multimodal therapy.

Objective To analyze the correlation of valproic acid and its metabolites (2-propyl-4-pentenoic acid, 3-hydroxy valproic acid,5-hydroxy valproic acid) with liver injury reference index. Methods 328 plasma samples from epilepsy patients were collected and divided into two groups（123 samples from patients with abnormal liver function, experimental group; 205 samples from patients with normal liver function, control group）.The plasma concentrations of valproic acid and its metabolites in the two groups were determined by LC-MS/MS method and the diagnostic value of the concentrations to liver disfunction was analyzed by ROC curve. Results The mean plasma concentration of valproic acid and its three metabolites in the patients with abnormal liver function was higher than that in the control group with was statistically difference（P<0.05）.The concentration of valproic acid and its metabolites could be used as a reference for the diagnosis of liver injury,5-hydroxy valproic acid had better diagnostic value than valproic acid. Conclusion The metabolites of valproic acid were associated with hepatotoxicity, which could be used as a diagnostic index of liver injury and could be a reference for clinical safe application of valproic acid.

Objective: To evaluate the risk factors of postoperative upgrade to pT₃a of cT₁ renal cell carcinoma, and to establish a nomogram prediction model to improve the ability of predicting locally advanced renal cell carcinoma and provide a reference for clinical surgical decision-making. Methods: Clinical data of 1 376 patients with cT₁ (diameter ≤ 7 cm) renal tumor hospitalized for surgery from January 2010 to December 2016 were retrospectively analyzed. There were 979 males and 397 females, with the mean age of (57.65±10.92) years. The mean body mass index (BMI) was (25.47±3.27) kg/m² and the average tumor size was (4.02±1.52) cm. There were 711 tumors on the left and 665 on the right. There were 363 cases with clinical symptoms, 567 patients with smoking history , 732 cases with history of chronic disease. There were 289 cases with tumor necrosis, 636 cases with tumor protrusion, 822 cases with irregular tumor , and 738 cases with renal sinus compression. Partial nephrectomy and radical nephrectomy were performed in 396 cases and 980 cases respectively. Mann-whitney U test and chi-square test were used for univariate analysis, logistic regression analysis was used for multivariate analysis to analyze the predictors of upgrading, R software was used to construct the nomogram predictive model, C-index was used to evaluate the model discrimination, and calibration curve method was used to evaluate the consistency of the model. Results: Postoperative pathology of total 1 376 cases showed that there were 1 195 cases of clear cell carcinoma of kidney, 48 cases of papillary cell carcinoma, 57 cases of chromophobe cell carcinoma, and 76 cases of other types. Among the 1 376 patients with cT₁ renal tumor, 75 patients were upgraded to pT_3a, accounting for 5.5% of all patients. Univariate analysis showed that the patients who upgraded to pT_3a were older [(63.08±10.17) years old and (57.34±10.88) years old], and the tumor length and diameter were larger [(5.24±1.35) cm and (3.95±1.51) cm]. Patients with clinical symptoms [46.7% (35/75) vs. 25.2%(328/1 301)], patients with CT indication of tumor necrosis [40.0%(30/25 975) vs. 19.9% (259/1 301)], patients with irregular tumor contour [73.3%(55/76 775) vs. 59.0%(767/1 301)], and patients with radical nephrectomy were higher [(92.1% (70/91 075) vs. 70%(910/1 301)]. All the differences were statistically significant (P<0.01). Multivariate analysis showed that the independent predictors of upgrading were age (OR=1.046, P<0.001), larger tumor (OR=1.504, P<0.001), clinically symptom (OR=2.153, P=0.004), irregular tumor profile (OR=2.466, P=0.002), and tumor necrosis on CT (OR=2.588, P<0.001). The C-index was 0.808, the calibration curve of forecasting curve with the standard curve fit was good, and the prediction of renal cancer are better in predict consistency. Conclusions Based on the five preoperative predictors, including age, tumor size, clinical presence or absence of symptoms, tumor profile, and whether or not the tumor necrosis indicated by CT, this study developed a nomogram of cT₁ renal cancer upgrade to pT_3a. This nomogram has a good statistical significance, and this model can provide prognosis consultation for patients and provide reference for doctors to make decisions before treatment.

OBJECTIVE: To investigate the long-term outcome of laparoscope-assisted transanal total mesorectal excision (taTME) for rectal cancer. METHODS: Clinicopathological data of 29 patients with mid-low rectal cancer undergoing laparoscope-assisted taTME at Department of Gastrointestinal Surgery, the First Affiliated Hospital of Guangzhou Medical University from May 2010 to December 2015 were collected for the retrospective case-series study. All the operations were performed with transabdominal and transanal procedure simultaneously or sequentially. Perioperative presentations, pathological examinations, and oncologic outcomes were retrospectively analyzed. Long-term recurrence, metastasis and survival were assessed during follow-up. Outpatient clinic and telephone survey were used for follow-up. The follow-up time ended in December 2018. The overall survival (OS) rate and disease-free survival (DFS) rate were calculated by the Kaplan-Meier method. RESULTS: The average intra-operative blood loss was (75.9±9.5) ml (range,20 to 200). The average operating time was (223.6±4.1) minutes (range, 165 to 280). The average number of harvested lymph node was 22.3±2.0. The average length of pathological specimen was (13.1±0.6) cm. The average distal resection margin was (2.9±0.2) cm. 89.7% (26/29) of specimens was complete and 10.3% (3/29) near complete. Two cases (6.9%) had positive cutting circumferential margin. Median follow-up was 56 (range, 22 to 91) months. Local recurrence rate, distant metastasis rate, 3-year OS rate, 3-year DFS rate, 5-year OS rate, 5-year DFS rate were 10.3% (3/29), 20.7%(6/29), 96.6%, 83.2%, 87.6% and 79.6%, respectively. No incisional hernia or adhesive intestinal obstruction was found. CONCLUSION Long-term outcomes of mid-low rectal cancer patients undergoing laparoscope-assisted taTME are satisfactory.
Humans ; Laparoscopes ; Neoplasm Recurrence, Local ; Rectal Neoplasms ; surgery ; Rectum ; Retrospective Studies ; Transanal Endoscopic Surgery

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.