Objective To find the effect of alter-expressed PER2 on proliferation,apoptosis and other clock genes expression in human oral squamous cell carcinoma SCC15 cells.Methods Short hairpin RNA interference was used to knockdown PER2 in SCC15 human oral squamous cell carcinoma cells.Flow cytometry analysis was used to testify the cell proliferation and apoptosis.Quantitative real-time PCR was used to testify the mRNA expressions of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα,NPAS2,PER1 and REV-ERBα.Results The proliferation was enhanced and apoptosis was decreased after PER2 knockdown in SCC15 cells (P<0.05).The mRNA expression of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα and NPAS2 was significantly down-regulated,and the mRNA expression of PER1 and REV-ERBα was significantly up-regulated (P<0.05).Conclusions Clock gene PER2 plays an important role in regulating other clock genes of the clock gene network in cancer cells,PER2 knockdown can enhance proliferation and recede apoptosis of cancer cell.

Objective To describe the development of nodules of altered hepatocytes (NAH) in chronic hepatitis B and to reveal progression of the nodules to hepatocellular carcinoma (HCC). Methods HCC, NAH and ordinary regenerative nodules (ORN) were identified and compared histologically. Expression levels of hepatitis B virus (HBV) antigens, mitoactivity and p53 accumulation in these lesions were evaluated by immunohistochemistry. Results Multiple foci of altered hepatocytes (FAH) and NAH were identified in the liver parenchyma surrounding HCC in all of the samples examined. Sequential architectural and cellular changes were observed during the progression of FAH to NAH and HCC. Expression levels of HBV surface and core antigens were found to be significantly decreased in ORN, NAH and HCC, with their positive rates being 70 % (35/50), 50 % (25/50), 10 % (5/50) and 60 % (30/50), 40 % (20/50), 6 % (3/50), respectively (P ＜0.05). Ki-67-1abelling indices were determined to be (0.58±0.49) %, (2.46±1.05) % and (40.36±26.27) %in these lesions, respectively (P ＜0.05). Nuclear p53 accumulation was found only in HCC. Its occurrence was associated to a high histological grade, with its frequencies being 13 % (1/8), 41% (11/27) and 73 % (11/15)in grade 1, 2 and 3 lesions, respectively. Conclusion NAH lesions, identified by their morphologic features and mitoactivity elevation, are detectable in resected liver samples with chronic hepatitis B and cirrhosis. They represent a common HCC precursor and can be used as a surrogate marker for the surveillance of high-risk individuals.

Objective Focal nodular hyperplasia(FNH) is composed of multiple hyperplastic liver cell nodules,but its pathogenesis has not been elucidated. Foci (FAH) or nodules of altered hepatocytes (NAH) are precursors of hepatocellular adenoma (HCA) and carcinoma.This study aimed at identifying FAH and NAH from FNH and evaluating their role in FNH development.Methods 6 FNH lesions from 5 patients and 10 HCA from 9 patients were examined histologically,and expression levels of CD_(34) cytokeratin 19(CKl9) and Ki-67 antigen were demonstrated immunohistochemicailly.Proliferative activity was evaluated by Ki-67 antigen-labeling indices(Ki-67 LI).Results Multiple FAH and NAH were identified in all of the 6 FNH lesions. Whiie micmvasculatures were demonstrated by CD_(34) immunoreactivity in both HCA and FNH,their density and distribution were different in these two lesions,being diffuse in HCA and focal or nodular,mainly within NAH.CKl9 expression Was found in FNH,localized in ductal and ductular cells,but not within NAH and HCA.Average Ki.67 LI of 73 NAH(2.8%) was shown to be higher than that of the whole FNH lesions (0.6%),and had no statistieal difference comparable to that of HCA(1.8%).Conclusion Muhiple NAH are present in all classical FNH lesions.Unlike the surrounding parenchyma,NAH lesions are more proliferative and equipped with CD_(34)-positive microvasculatures as in HCA.

OBJECTIVEThis study investigated the effect of clock gene PER1 on the expression levels of other clock genes in clock gene networks in oral squamous cell carcinoma cells.

METHODSWe used RNA interference mediated by short hairpin RNAs (shRNAs) to effectively knock down PER1 in SCC15 human oral squamous cell carcinoma cells. Flow cytometry was used to detect the degree of proliferation and apoptosis of the cells after PER1 knockdown, and quantitative real-time PCR was used to detect the mRNA expression levels of the clock genes CLOCK, BMAL1, PER1, PER2, PER3, DEC1, DEC2, CRY1, CRY2, TIM, CKIE, RORA, NPAS2, and REV-ERBA.

RESULTSThe proliferation index of SCC15 cells increased significantly while the apoptotic index decreased significantly after PER1 knockdown (P<0.05). The mRNA expression levels of PER1, PER2, DEC1, DEC2, CRY1, CRY2, and NPAS2 markedly decreased (P<0.05) while those of PER3, TIM, RORA, and REV-ERBA markedly increased (P<0.05). By contrast, no obvious changes were observed in the mRNA expression levels of CLOCK, BMAL1, and CKIE (P>0.05).

CONCLUSIONSThe clock gene PER1 can regulate the expression levels of other clock genes in the clock gene networks; these genes include PER2, DEC1, DEC2, CRY1, CRY2, NPAS2, PER3, TIM, RORA, and REV-ERBA. PER1 gene thus plays an important role in the regulation of clock gene networks.
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Objective:To explore the differences of wear resistance of three kinds of glass ceramics and Wieland Zenostar zircona (Zenostar) , and to clarify their influencing factors.Methods:Zenostar were made into flat-shaped specimens (zirconia base sample group) and hemisphere-shaped specimens (zirconia pair grinding group) .There kinds of glass ceramics IPS Empress (Empress) , IPS e.max CAD (e.max) , VITA Suprinity (Suprinty) were used as base specimens.Each group was exposed to UMT-2testing machine to simulate the clinical service.The wear depthes of base specimens were detected by laser confocal scanning.Scanning electron microscope (SEM) was used to evaluate the wear surfaces.Results:In zirconia base sample group, there were no significant differences in the maximum wear depthes to Zenostar between the three kinds of glass ceramics (P＞0.05) .In zirconia pair grinding group, the maximum wear depthes ranked as follows:Zenostar group＜e.max group≈Empress group＜Suprinity group;there was no significant difference between e.max group and Empress group (P＞0.05) , but there were significant differences between other groups (P＜0.01) .The SEM results showed that the wearing surface of the Zenostar in zirconia base sample group was relatively smooth;whereas the wearing surface of Empress in zirconia pair grinding group was rougher with alarge area of clebris desquamation surface.Conclusion:The wear resistance of the three kinds of glass ceramics to Zenostar is related to the compositions and the chemical structures of materials.

Objective:To investigate the effect of Astragaloside Ⅳ on high glucose-induced cardiomyocyte pyroptosis.Methods:H9c2 cells were cultured in vitro and divided into control group(5.5 mmol/L glucose), high glucose group(33.3 mmol/L glucose), Astragaloside Ⅳ group(33.3 mmol/L glucose+ 100μmol/L Astragaloside Ⅳ), and NLRP3 inhibitor group(33.3 mmol/L glucose+ 1μmol/L MCC950). Cell counting kit 8(CCK-8)was used to detect the activity of H9c2 cells.Lactate dehydrogenase(LDH)kit was used to detect the content of LDH in cell supernatant.Superoxide anion fluorescent probe(DHE)was used to detect the level of intracellular reactive oxygen species(ROS). Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression levels of pyroptosis-related genes.Immunofluorescence was used to detect the fluorescence intensity of NLRP3.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of inflammatory factors in cell supernatant.Results:When the concentration of Astragaloside Ⅳ was 100 μmol/L, it could significantly inhibit the decrease of cardiomyocyte viability induced by high glucose( P<0.01)and reduce LDH release( P<0.01). Compared with the control group, the level of ROS was increased( P<0.01), the mRNA and protein expressions of pyroptosis-related molecules were up-regulated( P<0.01 for all), the fluorescence intensity of NLRP3 was increased( P<0.01), and the levels of inflammatory factors in the cell supernatant were increased in the high glucose group( P<0.01). Compared with the high glucose group, the ROS level was decreased( P<0.01), the mRNA and protein expressions of pyroptosis-related molecules were down-regulated( P<0.05 or P<0.01), the fluorescence intensity of NLRP3 was decreased( P<0.01), and the levels of inflammatory factors in cell supernatant were decreased( P<0.05 or P<0.01)in Astragaloside Ⅳ group and inhibitor group. Conclusions:Astragaloside Ⅳ plays a protective role in high glucose-induced cardiomyocyte injury by inhibiting NLRP3/Caspase-1 signaling pathway and inhibiting pyroptosis.Moreover, it can improve the anti-inflammatory and antioxidant properties in cell models.

Objective: To develop the monoclonal antibody (mAb) against neuraminidase of H7N9 subtype influenza A virus and identify its biological function. Methods: Female 8 week-old BALB/c mice were immunized and the splenocytes of the mice were fused with Sp2/0 myeloma cells. Indirect ELISA was used to screen hybridoma and the positive clones were subject to be subcloned. Positive clones were identified and the monoclonal antibodies(mAbs) were obtained by purifying the ascetic fluid of mice injected with the hybridoma. The NA-binding as well as neuraminidase-inhibition activity of these mAbs were determined. Results: Three mAbs against neuraminidase of H7N9 subtype influenza A virus, 1G8, 3C4 and 4E8, were obtained. They demonstrated different epitop-recognizing. 3C4 and 4E8 exhibited neuraminidase inhibitory activity, with a IC₅₀ of 1.45 μg/ml and 8.65 μg/ml, respectively. Conclusions The results suggested that mAbs specific to neuraminidase of H7N9 subtype influenza A virus were developed, providing an useful tool in control and preventing the novel H7N9 influenza A virus.

Objective: To develop a model incorporating radiomic features and clinical factors to accurately predict acute ischemic stroke (AIS) outcomes. Materials and Methods: Data from 522 AIS patients (382 male [73.2%]; mean age ± standard deviation, 58.9 ± 11.5 years) were randomly divided into the training (n = 311) and validation cohorts (n = 211). According to the modified Rankin Scale (mRS) at 6 months after hospital discharge, prognosis was dichotomized into good (mRS ≤ 2) and poor (mRS > 2); 1310 radiomics features were extracted from diffusion-weighted imaging and apparent diffusion coefficient maps. The minimum redundancy maximum relevance algorithm and the least absolute shrinkage and selection operator logistic regression method were implemented to select the features and establish a radiomics model. Univariable and multivariable logistic regression analyses were performed to identify the clinical factors and construct a clinical model. Ultimately, a multivariable logistic regression analysis incorporating independent clinical factors and radiomics score was implemented to establish the final combined prediction model using a backward step-down selection procedure, and a clinical-radiomics nomogram was developed. The models were evaluated using calibration, receiver operating characteristic (ROC), and decision curve analyses. Results: Age, sex, stroke history, diabetes, baseline mRS, baseline National Institutes of Health Stroke Scale score, and radiomics score were independent predictors of AIS outcomes. The area under the ROC curve of the clinical-radiomics model was 0.868 (95% confidence interval, 0.825–0.910) in the training cohort and 0.890 (0.844–0.936) in the validation cohort, which was significantly larger than that of the clinical or radiomics models. The clinical radiomics nomogram was well calibrated (p > 0.05). The decision curve analysis indicated its clinical usefulness. Conclusion The clinical-radiomics model outperformed individual clinical or radiomics models and achieved satisfactory performance in predicting AIS outcomes.

OBJECTIVE To comprehensively screen the optimal steaming time of salt-steaming Morinda officinalis （SSMO） based on Q-markers and anti-oxidative activities， and to establish characteristic quality standard of the decoction pieces. METHODS The contents of six Q-markers （1-kestose， nystose， 1F-fructofuranosylnystose， inulotriose， inulotetraose and inulopentaose） in SSMO at different steaming time were determined by HPLC-ELSD method simultaneously. The activity of sample extracts to scavenge 4 kinds of oxidative free radical and their iron reduction abilities were determined by visible UV spectrophotometer. The optimal steaming time of SSMO was screened by gray relevance degree and entropy weight technique for order preference by similarity to an ideal solution （TOPSIS）-fusion model method. The contents of six Q-markers in 10 batches of SSMO prepared at the optimal steaming time were determined. HPLC-ELSD fingerprints of SSMO decoction pieces were established. RESULTS The results showed that the contents of six Q-markers were the highest when SSMO was steamed for 3-5 h； and the ability of scavenging DPPH·， ABTS·， PTIO·， ·OH and iron reduction ability was the best at 5 h. There were 20 common peaks in the fingerprints for 10 batches of samples， and the similarities were higher than 0.990. A total of 9 chromatographic peaks were identified， which were D-fructose （peak 1）， D（+）-glucose （peak 2）， sucrose （peak 3）， 1-kestose （peak 4）， nystose （peak 5）， 1F-fructofuranosylnystose （peak 6）， inulotriose （peak X2）， inulotetraose （peak X3） and inulopentaose （peak X4）. Average contents of six Q-markers were 4.17%， 5.54%， 6.60%， 2.89%， 2.62% and 2.13%， respectively. CONCLUSIONS The optimal steaming time of SSMO is 5 h； the contents of six Q-markers are primarily determined on the basis of dry product， which are no less than 3.03%， 4.11%， 4.87%， 2.15%， 1.96% and 1.58%， respectively. The ratio of Inulin-/Inulo oligosaccharides content is no more than 2.5.

Objective To investigate the epidemiological characteristics of foodborne diseases in Liuzhou City, and to provide reference for formulating the prevention and control measures of foodborne diseases. Methods The surveillance data of foodborne diseases in 25 sentinel hospitals in Liuzhou City from 2018 to 2020 were collected for statistical analysis. Results From 2018 to 2020, a total of 9 317 cases of foodborne diseases were reported in Liuzhou City, and 2 158 samples were collected for pathogen detection. A total of 311 cases were detected positive , with a detection rate of 14.41%. Norovirus had the highest detection rate (8.63%), followed by Salmonella (4.08%) and Escherichia coli (3.10%). July to October was a period of high incidence of foodborne disease（ 41.17%）. The proportion of patients aged 60 and over was the highest (18.49%), followed by the age group of 30-39 (18.03%). Suspicious foods were mainly meat and meat products (22.35%) and aquatic animals and their products (13.89%). The suspicious eating places were mainly families (40.43%) and restaurants (13.63%). Conclusion The high incidence of foodborne diseases in Liuzhou occurs in summer and autumn. The main pathogens are Salmonella and norovirus. Infected patients are concentrated in the age group of 60 years and above and the age group of 30 to 39 years old. The family is the main place of foodborne disease, followed by the restaurants and hotels. Suspicious foods include mainly meat and meat products and aquatic animals and their products. It is necessary to strengthen monitoring ability and food safety education to reduce the occurrence of foodborne diseases.