[Effect of Shikonin on Autophagy and Apoptosis of Human Promyelocytic Leukemia Cells].

OBJECTIVE: To explore the effect of shikonin on autophagy and apoptosis of human promyelocytic leukemia cells and its possible mechanism. METHODS: Human promyelocytic leukemia cells NB4 in the logarithmic growth phase were divided into control group (untreated NB4 cells), shikonin group (0.3 µmol/L shikonin treatment), 740Y-P group (15 µmol/L PI3K/Akt/mTOR pathway activator 740Y-P treatment), shikonin+740Y-P group (0.3 µmol/L shikonin and 15 µmol/L 740Y-P co-treatment), after 24 hours of treatment, the cells were used for subsequent experiments. CCK-8 method was used to detect cell viability, monodansylcadaverine (MDC) staining to detect the aggregation of autophagic vesicles, flow cytometry to detect cell apoptosis, and Western blot to detect the expression of Beclin1, LC3, p62, Bax, cleaved caspase-3, Bcl-2 and PI3K/Akt/mTOR pathway related proteins. RESULTS: Compared with the control group, the purple punctate fluorescence intensity, apoptosis rate, Beclin1, LC3-Ⅱ/LC3-Ⅰ, cleaved caspase-3, and Bax protein expression in NB4 cells were increased in the shikonin group, while OD450 value (24, 48 h) and the expressions of Bcl-2 and p62 proteins were decreased (all P < 0.05). Compared with the control group, the purple punctate fluorescence intensity, apoptosis rate, Beclin1, LC3-Ⅱ/LC3-Ⅰ, cleaved caspase-3, and Bax protein expression in NB4 cells were decreased, while OD450 value (24, 48 h) and the expressions of Bcl-2 and p62 proteins were increased in the 740Y-P group (all P < 0.05). Compared with the shikonin group, the purple punctate fluorescence intensity, apoptosis rate, Beclin1, LC3-Ⅱ/LC3-Ⅰ, cleaved caspase-3, and Bax protein expression in NB4 cells were decreased, while OD450 value (24, 48 h) and the expressions of Bcl-2 and p62 proteins were increased in the shikonin+740Y-P group (all P < 0.05). Compared with the control group, the expression of PI3K/Akt/mTOR pathway related proteins p-PI3K, p-Akt, and p-mTOR in NB4 cells were significantly decreased in the shikonin group, while those in the 740Y-P group were increased (all P < 0.05). Compared with the shikonin group, the expressions of p-PI3K, p-Akt, and p-mTOR proteins in NB4 cells were significantly increased in the shikonin+740Y-P group (all P < 0.05). CONCLUSION: Shikonin may promote autophagy and apoptosis of NB4 cells by inhibiting PI3K/Akt/mTOR pathway.

Assessment tools for stigma in breast cancer patients based on COSMIN guidelines: a systematic review.

OBJECTIVE: The objective of this study was to conduct a systematic review of the measurement properties and methodological quality of stigma assessment tools designed for breast cancer patients. The aim was to provide clinical medical staff with a foundation for selecting high-quality assessment tools. METHODS: A comprehensive computer search was carried out across various databases, including SinoMed, China National Knowledge Infrastructure (CNKI), Wanfang Database, China Science and Technology Journal Database(VIP), Embase, PubMed, Web of Science, The Cochrane Library, and Scopus, which were searched from the inception of the databases until March 20, 2023. Literature screening and data extraction were performed independently by two researchers, adhering to predefined inclusion and exclusion criteria. The assessment tools were evaluated using the Consensus-based Standards for the selection of health Measurement Instruments (COSMIN) systematic evaluation guidelines. RESULTS: In the final analysis, a total of 9 assessment tools were included. However, none of these tools addressed measurement error, cross-cultural validity, criterion validity, and responsiveness. Following the COSMIN guidelines, BCSS and CSPDS were assigned to Class A recommendations, while the remaining tools received Class B recommendations. CONCLUSION: The BCSS and CSPDS scales demonstrated comprehensive assessment in terms of their measurement characteristics, exhibiting good methodological quality, measurement attribute quality, and supporting evidence. Therefore, it is recommended to utilize these scales for evaluating breast cancer stigma. However, further validation is required for the remaining assessment tools.

[Effect of three mixing methods on air bubbles and solidification state of alginate impression materials].

PURPOSE: To evaluate the effect of full-automatic mixing machine method, clockwise manual mixing and combined eight-shaped manual mixing on air bubble content, flowability, temperature, working time and setting time of alginate impression materials. METHODS: With the same condition, alginate impression materials were mixed by three different methods. The number of bubbles, area, flowability, temperature, working time and setting time were evaluated with SPSS 24.0 software package. RESULTS: The number of bubbles in the automatic mixing group was (2.30±2.50), and the area was (0.17±0.18) mm2, which was less than the number of clockwise manual mixing group (59.60±14.19), and the total area (7.41±2.24) mm2 (P＜0.01). The flowability of the clockwise manual mixing group [(39.52±0.85) mm] was less than that of the full-automatic mixing group [(50.78±0.90) mm] and the combined eight-character manual mixing group [(50.36±1.75) mm](P＜0.01).The setting time of the material mixed by three methods was eligible for clinical use. CONCLUSIONS: The mixing method of alginate impression material has an effect on material's bubble content, flowability and temperature changes. The impression materials mixed by full-automatic mixing method are better in terms of bubble content, flowability and other properties. If manual mixing is used, combined eight-shaped manual mixing method can help reduce impression bubbles and deformation, and improve flowability.

[Relationship between sICAM-1, IL-1ß and HIF-1α levels in gingival sulcus fluid and peri-implantitis in patients with implant restoration].

PURPOSE: To investigate the relationship between the levels of soluble intercellular adhesion molecule-1 (sICAM-1), interleukin-1ß (IL-1ß) and hypoxia-inducible factor-1α (HIF-1α) in gingival sulcus fluid and peri-implantitis (PI) in patients with implant restoration. METHODS: A total of 198 patients with implant restoration admitted to Fengcheng Hospital from January 2019 to December 2021 were selected, the patients were divided into PI and non-PI group according to whether the implant restoration was complicated by PI 3 months after restoration. The levels of sICAM-1, IL-1ß and HIF-1α in the gingival sulcus fluid prior to implant restoration were measured by enzyme-linked immunosorbent assay. Multi-factor logistic regression was used to analyze the factors influencing concurrent PI in patients with implant restoration. ROC curves were used to analyze the predictive value of sICAM-1, IL-1ß and HIF-1α levels in gingival sulcus fluid on concurrent PI in patients with implant restoration. SPSS 28.0 software package was used for statistical processing of the data. RESULTS: The incidence of PI in 198 patients with implant restoration was 17.68% (35/198) 3 months after implant restoration. The levels of sICAM-1, IL-1ß and HIF-1α in the gingival sulcus fluid were significantly higher in the PI group than in the non-PI group (P＜0.05). Multi-factor logistic regression analysis showed that elevated sICAM-1(OR=1.135, 95%CI: 1.066-1.208), IL-1ß (OR=1.106, 95%CI: 1.054-1.161) and HIF-1α (OR=1.008, 95%CI: 1.004-1.012) were independent risk factors for complications of PI in prosthetic patients(P＜0.05). ROC curve analysis showed that the area under the curve for sICAM-1, IL-1ß and HIF-1α levels in gingival sulcus fluid alone and in combination for the diagnosis of concurrent PI in patients with implantation was 0.787, 0.785, 0.794 and 0.930, respectively, with sensitivity of 80.00%, 74.29%, 62.86% and 88.57% and specificity of 66.87%, 74.85%, 78.53% and 85.28%, respectively. CONCLUSIONS: Elevated levels of sICAM-1, IL-1ß and HIF-1α in gingival sulcus fluid are independent risk factors for PI complications in patients with implant restoration and can be used as an auxiliary predictor of PI complications in patients with implant restoration.

LRP6 Bidirectionally Regulates Insulin Sensitivity through Insulin Receptor and S6K Signaling in Rats with CG-IUGR.

OBJECTIVE: Intrauterine growth restriction followed by postnatal catch-up growth (CG-IUGR) increases the risk of insulin resistance-related diseases. Low-density lipoprotein receptor-related protein 6 (LRP6) plays a substantial role in glucose metabolism. However, whether LRP6 is involved in the insulin resistance of CG-IUGR is unclear. This study aimed to explore the role of LRP6 in insulin signaling in response to CG-IUGR. METHODS: The CG-IUGR rat model was established via a maternal gestational nutritional restriction followed by postnatal litter size reduction. The mRNA and protein expression of the components in the insulin pathway, LRP6/ß-catenin and mammalian target of rapamycin (mTOR)/S6 kinase (S6K) signaling, was determined. Liver tissues were immunostained for the expression of LRP6 and ß-catenin. LRP6 was overexpressed or silenced in primary hepatocytes to explore its role in insulin signaling. RESULTS: Compared with the control rats, CG-IUGR rats showed higher homeostasis model assessment for insulin resistance (HOMA-IR) index and fasting insulin level, decreased insulin signaling, reduced mTOR/S6K/ insulin receptor substrate-1 (IRS-1) serine307 activity, and decreased LRP6/ß-catenin in the liver tissue. The knockdown of LRP6 in hepatocytes from appropriate-for-gestational-age (AGA) rats led to reductions in insulin receptor (IR) signaling and mTOR/S6K/IRS-1 serine307 activity. In contrast, LRP6 overexpression in hepatocytes of CG-IUGR rats resulted in elevated IR signaling and mTOR/S6K/IRS-1 serine307 activity. CONCLUSION: LRP6 regulated the insulin signaling in the CG-IUGR rats via two distinct pathways, IR and mTOR-S6K signaling. LRP6 may be a potential therapeutic target for insulin resistance in CG-IUGR individuals.

[A Review of Chronic Stress and the Initiation and Evolution of Cancer].

Chronic stress activates the typical neuroendocrine system, hypothalamus pituitary adrenal axis and sympathetic nervous system, and leads to a sustained non-specific adaptive response. It has been proved that chronic stress can promote tumor initiation and induce tumor evolution, especially in immune function and remodeling of tumor microenvironment. However, due to the complex mechanism of chronic stress and the great difference in individual tolerance, the research evidence of chronic stress in tumor genesis and progression is still unclear. Therefore, in this paper, we review the research on the relationship between chronic stress and tumor initiation and evolution, focusing on the molecular mechanism of chronic stress promoting tumor occurrence and development, inhibiting immune response and remodeling tumor immune microenvironment, and exploring the stress management program of healthy people and cancer patients, so as to provide clues for exploring new strategies of cancer prevention and treatment. In our opinion, targeting the cAMP/PKA/CREB signaling pathway to reverse tumor treatment strategy, the relationship between the tumor and stress, inflammation, immunity, the suppressor activity of ß receptor antagonist and its mechanism as well as associated with different treatment options, still need to be further explored. A healthy lifestyle, positive life attitudes and professional stress management guidance are essential for the prevention and treatment of cancer.

[Clinical analysis of muscle function intensifier in treatment of 40 patients with maxillary protrusion].

PURPOSE: To investigate the clinical effects of combined muscle functions intensifier and straight wire appliance in the treatment of maxillary protrusion. METHODS: Forty patients with maxillary protrusion were randomly divided into 2 groups (group A and group B). Straight wire appliance and muscle function intensifier were used in group A, while patients in group B received straight wire appliance only. X-ray projection was conducted to measure relative maxillary index (labial inclination of upper or lower incisor, SNB angle, ANB angle, FMA, maxillary plane angle, NA-PA, U1-FH, U1-PP, forward moving distance of U6) before and 6 months after treatment. The efficacy was analyzed and evaluated using SPSS 20.0 software package. RESULTS: After correction, labial inclination of upper or lower incisor, SNB angle, ANB angle, FMA, maxillary plane angle, NA-PA, U1-FH, U1-PP changed significantly in both groups (P<0.05), but patients in group A achieved more incisor adduction and less forward moving distance of U6, with significant difference between the two groups (P<0.05). CONCLUSIONS: Most patients with maxillary protrusion have a bad habit of mouth breathing. Muscle function intensifier is used only at night and can relieve the patient's mouth breathing, speed up the movement of teeth, and reduce the loss of anchorage and ultimately, improve the correction efficiency and clinical effect of patients with maxillary protrusion.

Aberrant hypermethylation and reduced expression of disabled-2 promote the development of lung cancers.

Disabled-2 (Dab2) is considered a tumor suppressor and is downregulated in cancers. We examined the promoter methylation status and expression levels of Dab2, and investigated their roles in the development of lung cancers. Methylation-specific PCR was employed to analyze the methylation status of Dab2 in 100 lung cancer tissues. The cytoplasmic and nuclear expression of the Dab2 protein was determined using western blot analysis. Demethylation treatment using 5-Aza-2-deoxycytidine (5-Aza-dC) was performed in three lung cancer cell lines. Dab2 expression was upregulated by Dab2 transfection or interrupted by Dab2 siRNA in lung cancer cells. Proliferative and invasive ability tests were performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTS) and a Matrigel invasion assay, respectively. The methylation rate of Dab2 was significantly higher in lung cancer tissues compared to normal lung tissues. Dab2 methylation correlated with the reduced nuclear and cytoplasmic expression of Dab2, as well as the TNM stage and lymphatic metastasis of lung cancers. Treatment with 5-Aza-dC was able to eliminate the hypermethylation of Dab2, enhance Dab2 expression, and inhibit ß-catenin expression, and the proliferative and invasive ability of lung cancer cells. Upregulation of Dab2 expression reduced ß-catenin expression and proliferation and invasiveness of lung cancer cells. However, interruption of Dab2 expression induced the opposite results. Dab2 methylation is common in lung cancers, and is one of the most important factors responsible for the reduced expression of Dab2. Aberrant hypermethylation and reduced expression of Dab2 promote the development of lung cancers.

[Current situation on the China infectious disease automated-alert and response system in Guangdong Province, China].

OBJECTIVE: To analyze the current situation on China Infectious Disease Automated-alert and Response System (CIDARS) in Guangdong province, China. METHODS: Early-warning signals and response time were analyzed by using three approaches of CIDARS. Positive rates of early-warning signals and error early-warning rates prior and after the adjustment,were compared. RESULTS: Totally, 114 585 early-warning signals appeared, with an average response time of 1.35 hours from April 21, 2008 to December 31, 2012. There were 12,394 early-warning signals in terms of fixed threshold method with a positive rate of 7.96%. 85 727 early-warning signals appeared under the mobile percentile method with a positive rate of 0.85%. There were 16,464 early-warning signals by using accumulation and control chart methods,with a positive rate of 1.82%. Results showed there was a positive correlation between the number of reported cases and the number of early-warning signals (r = 0.924, P < 0.01). The overall positive rate in Guangdong province increased from 1.48% to 2.14%, after the adjustment done by the Chinese Center for Disease Control and Prevention in 2010. The error early-warning rates regarding eight infectious diseases including hepatitis A, bacillary or amebic dysentery, both typhoid and paratyphoid fevers had reduced. CONCLUSION: The early-warning signal response appeared timely in CIDARS with good operation. However, despite the improvement on the efficacy of CIDARS, some functions and parameters of the systems still need to be adjusted.

Atonal homolog 1 expression in lung cancer correlates with inhibitors of the Wnt pathway as well as the differentiation and primary tumor stage.

Atonal homolog 1 (Atoh1) is crucial to the differentiation of many cell types and participates in tumorigenesis and progression. This study investigated the role of Atoh1 in lung cancer development and its correlation with key members of the Wnt pathway. We used immunohistochemistry to examine the expressions of Atoh1, ß-catenin, Axin, chibby, and Disabled-2 (Dab2) in 118 samples of lung cancer. We also detected the cytoplasmic and nuclear expression of Atoh1 in lung cancer tissues using western blot. Atoh1 nuclear expression was negatively correlated with differentiation level (p = 0.004) and primary tumor stage (p = 0.044) of lung cancer. Nuclear Atoh1 expression was positively correlated with nuclear expression of chibby (p < 0.001) and Dab2 (p < 0.001). Cytoplasmic Atoh1 expression was positively correlated with the cytoplasmic expression of Axin (p = 0.028), chibby (p < 0.001), and Dab2 (p < 0.001). We conclude that the nuclear expression of Atoh1 was inversely correlated with the differentiation and primary tumor stage of lung cancers. The expression and localization of Atoh1 correlated with Axin, chibby, or Dab2. Atoh1 may be a potential therapeutic target for the inhibition of growth and progression of lung cancers.

The expression patterns and correlations of chibby, ß-catenin, and DNA methyltransferase-1 and their clinicopathological significance in lung cancers.

Chibby is an inhibitor of the Wnt pathway. The expression and correlation of chibby in lung cancers is unclear. We considered that the expression pattern of chibby might be related to the expression of ß-catenin and DNA methyltransferase-1 (DNMT1). We examined the expressions of chibby, ß-catenin, and DNMT1 in 85 lung cancer tissues and corresponding normal lung tissues using immunohistochemistry. The nuclear expression rate of chibby was reduced in lung cancers (p < 0.001). Increased expression of DNMT1 was correlated with the differentiation (p = 0.034) and TNM stage (p = 0.048). The cytoplasmic expression of ß-catenin was correlated with poor differentiation of lung cancers (p = 0.016). The cytoplasmic and membrane expression of chibby was positively correlated with the cytoplasmic (p = 0.025) and membrane (p = 0.029) expressions of ß-catenin. The membrane expression of chibby was negatively correlated with the expression of DNMT1 (p = 0.006). Moreover, the expression of DNMT1 was correlated with the cytoplasmic expression of ß-catenin (p < 0.001). The nuclear expression of chibby is reduced in lung cancers. Chibby may colocalize with ß-catenin. ß-Catenin and DNMT1 may be concurrently expressed and thereby promote the development of lung cancers.

Disabled-2 and Axin are concurrently colocalized and underexpressed in lung cancers.

Disabled-2 expression is reduced in many cancers, suggesting that it is a potential tumor suppressor protein. To elucidate the role of Disabled-2 in lung cancer, we examined the expression of Disabled-2, the Disabled-2-binding protein Axin, and DNA methyltransferase-1 in lung cancer tissues and corresponding normal lung tissues using immunohistochemistry and Western blots. We also determined the subcellular localization of Axin and Disabled-2 in A549 cells using confocal immunofluorescence. Disabled-2 expression was significantly reduced in lung cancers and was colocalized and coexpressed with Axin (correlation coefficient = 0.321, P < .001 for cytoplasmic expression; correlation coefficient = 0.393, P < .001 for nuclear expression). Reduced nuclear Disabled-2 expression was correlated with the differentiation (P = .048) and TNM stage (P = .048) of the tumor. The cytoplasmic expression of Axin was also correlated with differentiation (P = .042), whereas the nuclear expression of Axin was correlated with both histologic type (P = .001) and TNM stage (P < .001) of lung cancers. Expression of DNA methyltransferase-1 was negatively correlated with the cytoplasmic expression of Axin (correlation coefficient = -0.244, P = .012) but positively correlated with the histologic type (P = .004), differentiation (P = .036), TNM stage (P = .044), and lymphatic metastasis (P = .011). Expressions of Disabled-2 and Axin were concurrently reduced and correlated with the malignant phenotype of lung cancers. Enhanced expression of DNA methyltransferase-1 correlated with the reduced expression of Axin and could be a marker for lung cancer development and progression.

[Clinical application of self-regulating exterior bracket for zygomatic fracture].

PURPOSE: To investigate the clinical value of self-regulating exterior bracket for zygomatic fracture. METHODS: Thirty patients with unilateral zygomatic fracture were treated using self-regulating exterior bracket. They all had limitation of mouth opening and facial collapse. 1.5 cm incision was made from the distal upper first molar. After reducting to normal mouth opening and symmetrical face, self-regulating external bracket was used to fix the fracture for 3 weeks. RESULTS: After removing the self-regulating bracket, X-ray showed the fracture healing well. The patients had normal mouth opening. CONCLUSION: The clinical effect of using self-regulating bracket to fix fracture after reduction for unilateral zygomatic fracture is good, it is a simple surgical method.

[Clinical study of two fixed prosthetics in dentition defect of Kennedy I and II type].

PURPOSE: To investigate the feasibility and clinical effect of two fixed prosthetic, supported by implant alone or by implant and natural teeth, in dentition defect of Kennedy I or II type. METHODS: Sixty subjects with dentition defect of Kennedy I or II were collected. They were divided into group A and group B, 30 subjects in each group respectively. Group A was restore with PFM crown or bridge, supported by implant or implant combined with natural teeth. Group B was treated by cantilever fixed bridge supported by natural teeth. The treatment outcomes were analyzed with SPSS17.0 software package. RESULTS: With 5 years of follow-up, all subjects in group A and 26/30 subjects in group B had successful restoration. There was significant difference in successful rate between group A and group B(P<0.05). CONCLUSION: Fixed prosthetics supported by implant alone and by both implant and natural teeth, in dentition defect of Kennedy I or II is feasible and its clinical effect is satisfactory. Supported by Research Fund of Science and Technology Commission of Fengxian District, Shanghai Municipality(Grant No.96-3).

[Clinical stability of immediate load of domestic micro-titanium screws for anchorage in orthodontics].

PURPOSE: To investigate the immediate load stability of domestic micro-titanium screws for anchorage in orthodontics. METHODS: Adult orthodontic patients needing the greatest anchorage were included in the study. Domestic micro-titanium screws were used to be orthodontic anchorage, immediate implantation and loading were conducted. RESULTS: A total of 52 implants of domestic micro-titanium anchorage in 20 patients were used. Forty-seven implants were successful in 20 patients,5 failed. The total loading time was 3-8 months. CONCLUSIONS: Domestic micro-titanium implant screws for anchorage can meet the needs of clinical orthodontics. However, immediate loading results in 10.4% failure rate, especially for the implant with a diameter of 1.2mmx7mm. Supported by Science and Technology Development Foundation of Fengxian District, Shanghai Municipality (Grant No. 090602).

[Clinical report of early implant-supported mandibular overdenture retained by conical crowns].

PURPOSE: To investigate the feasibility and clinical effectiveness of inchoate implant-supported mandibular overdenture retained by conical crowns. METHODS: In this study, 25 patients with edentulous mandible were included.All patients received 4 to 6 dental implants between the mental foramen and impression taking immediately to make overdenture retained by conical crowns,loading in ten days.All patients were followed up for 1,3,6,12,24 and 36 months, and then once a year.The stability of one implants, the state of peri-implant soft-tissue were recorded, and calculus of peri-implant was cleared. And the state of marginal bone resorption, osseointegration of implants were examined by X-ray films. RESULTS: A total of 103 implants were loaded early, 3 failed.The others had osseointegration,no infection of peri-implant soft-tissue occurred,no implants exhibited peri-implant radiolucencies. The patients were satisfied with the outcomes. CONCLUSIONS: The results of this clinical report suggest that immediate loading of the implants by conical crowns to support overdenture prostheses for edentulous mandible is feasible between the mental foramen. The short-term clinical effectiveness of early loading is good.