Objective As a signaling molecule,NO regulates key physiological processes and is closely related to periodontitis.To investigate the effect of flavonoid NO donor composite nanoparticles(G10@HAP/MSN@ZnO@COS)on osteogenic differentiation of periodontal ligament stem cells(PDLSCs)by regulating macrophage polarization.Methods The novel NO donor drug G10 was loaded on hydroxyapatite/mesoporous silicanant particles(HAP/MSN),filled with zinc oxide(ZnO),and then coated with chitosan(COS)to prepare composite nanoparticles(G10@HAP/MSN@ZnO@COS).The best concentration of G10@HAP/MSN@ZnO@COS was screened to promote cell proliferation by CCK-8 cell experiment.After the mouse mononuclear macrophages were stimulated by lipopo-lysaccharide,the mice were divided into four groups:Control group,G10 group,HAP/MSN@ZnO@COS group and G10@HAP/MSN@ZnO@COS group.Each group was cultured with fresh medium,5 μg/mL G10,5 μg/mL HAP/MSN@ZnO@COS and 5 μg/mL G10@HAP/MSN@ZnO@COS for 72 h respectively.ELISA and RT-qPCR were used to detect the expression of cytokines(TNF-α,IL-6,IL-1β,iNOS,IL-10)and mRNA expression in each group,and the phenotypic changes of M1/M2 were evaluated.The supernatant of each culture medium was used as conditioned medium to culture PDLSCs,and the osteogenic ability and cell miner-alization were evaluated by alkaline phosphatase activity test and alizarin red staining.Results CCK-8 experiment showed that G10@HAP/MSN@ZnO@COS of 5 μg/mL could significantly promote the proliferation of PDLSCs.The results of ELISA showed that compared with Control group,the expression of M1 type marker IL-1β,IL-6,TNF-α and iNOS in G10@HAP/MSN@ZnO@COS group was significantly decreased(P＜0.000 1),while the expression of M2 type marker IL-10 was significantly increased(P＜0.000 1).The results of RT-qPCR were consistent with those of ELISA,which showed that the expression of M1-related genes in G10@HAP/MSN@ZnO@COS group decreased significantly(P＜0.01).The results of alizarin red staining and alkaline phosphatase activity test showed that the number of mineralized nodules and alkaline phosphatase activity in G10@HAP/MSN@ZnO@COS-CM group were significantly higher than those in other groups(P＜0.000 1).Conclusion Composite nanoparticles(G10@HAP/MSN@ZnO@COS)can effectively inhibit the polarization of macrophages to M1 phenotype and promote it to M2 phenotypic polarization.The anti-inflammatory microenvironment regulated by G10@HAP/MSN@ZnO@COS can en-hance the osteogenic differentiation of PDLSCs.

Objective: To analyze the trend of lung cancer incidence rate among rural residents in Feicheng city between the years 2000 and 2012,and predict the incidence rate between the years 2013 and 2018,and subsequently provide baseline data for lung can-cer control and prevention.Methods:With the cancer registration data in Feicheng rural areas,the time trend of lung cancer inci-dence rate and the annual percentage change(APC)were calculated by the Joinpoint model,while the incidence of lung cancer from 2013 to 2018 were predicted by the ARIMA(p,d,q)model.Results:A total of 3,908 new cases of lung cancer were diagnosed be-tween 2000 to 2012.The incidence rate was 40.77/105,the age-specific cancer incidence rate in the Chinese population(ASRC)and world population(ASRW)were 32.95/105 and 32.97/105,respectively.The incidence was 2.14 times higher among males than females (P<0.001).The incidence of lung cancer which apparently rose from 25.13/105to 64.92/105 with an APC value of 9.74%(P<0.001),was increasing every year.The change in the trend of lung cancer with respect to age could be divided into three segments,the incidence rate in the 0 to 59 years group showed a rapid upward trend(APC=113.38,P<0.001),which was lower in the 60 to 79 years group (APC=20.39,P<0.05)and began to decline in the 80 years or older group(APC=-21.20,P>0.05).The incidence of lung cancer was also observed to be increasing yearly from 2013 to 2018,and with an average annual growth rate of 4.92%,was predicted to reach 87.92/105 in 2018.Conclusions:The occurrence of lung cancer was closely related to population aging,unhealthy habits,and environmental risk factors.Due to the increasing aging population,the incidence of lung cancer will continue to increase.In order to formulate specif-ic strategies,the control and prevention of lung cancer must be based on its incidence features.

Objective:To study the clinical characteristics of patients with brucella bloodstream infection in different age groups, and provide a basis for clinicians to take targeted diagnosis and treatment measures. Methods:Demographic data and general condition (age, sex, occupation, location, onset season, source of infection, clinical stage), clinical characteristics (main clinical symptoms and complications), and laboratory test results (routine and pathogenic tests) of adult patients with brucella bloodstream infection admitted to the Affiliated Hospital of Chengde Medical College from January 2015 to January 2020 were collected. According to the age stratification standards recommended by the World Health Organization, the patients were divided into a young group (18 - 44 years old), a middle-aged group (45 - 59 years old), and an elderly group (≥60 years old), and various indicators among different age groups were compared and analyzed. Results:A total of 75 patients were included, including 15 cases (20.00%) in the young group, 37 cases (49.33%) in the middle-aged group, and 23 cases (30.67%) in the elderly group. Among them, 61 cases (81.33%) were males and 14 cases (18.67%) were females, with statistically significant differences in gender ratios among different age groups (χ 2 = 7.28, P = 0.021). The majority of patients were farmers (64 cases, 85.33%), and 92.00% (69/75) of the patients came from rural areas. The main sources of infection were infected cattle and sheep, and contaminated food (39 cases, 52.00%). The main season of onset was spring and summer (45 cases, 60.00%). The clinical staging was mainly in the acute phase (66 cases, 88.00%). In terms of clinical symptoms, the young group of patients had no symptoms of low back pain, while the incidence rates of low back pain in the middle-aged and elderly groups were 35.14% (13/37) and 30.43% (7/23), respectively. There was a statistically significant difference between the three groups (χ 2 = 6.98, P = 0.031). In terms of complications, there were no cases of concurrent spondylitis in the young group of patients. The incidence rates of spondylitis in the middle-aged and elderly groups were 32.43% (12/37) and 34.78% (8/23), respectively. There was a statistically significant difference among the three groups (χ 2 = 6.86, P = 0.032). In terms of routine laboratory examinations, there were statistically significant differences in the proportion of blood lymphocytes and albumin levels among patients of different age groups ( F = 3.41, 3.27, P = 0.038, 0.044). In terms of pathogenic examination, there was a statistically significant difference in the median alarm time for positive blood culture among patients of different age groups ( H = 9.54, P = 0.008), with the middle-aged group having the longest (66.24 h) and the elderly group having the shortest (58.80 h). Conclusions:The clinical characteristics of patients with brucella bloodstream infection vary among different age groups, middle-aged and elderly patients are prone to low back pain symptoms, accompanied by spondylitis. Clinicians should pay attention to the patient's own characteristics and provide targeted diagnosis and treatment.

Objective To study the molecular mechanism of cisplatin to enhance the ability of tumor necrosis factor?related apoptosis?inducing ligand ( TRAIL ) in reversing multidrug resistance in vincristine?resistant human gastric cancer SGC7901/VCR cells. Methods MTT assay was used to measure the 50% inhibiting concentration( IC50 ) and cell survival in SGC7901 and SGC7901/VCR cells after different treatments.SGC7901/VCR cells were treated with different concentrations of DDP, different concentrations of TRAIL alone or in combination, and then the mRNA and protein levels of several genes were determined by RT?PCR, RT?qPCR and Western?blot analysis. After targeted silencing with specific siRNA and transfection of recombinant plasmid c?myc into the SGC7901/VCR cells, the mRNA and protein levels of DR4, DR5 and c?myc were determined by RT?PCR and Western?blot analysis. Results After combined treatment with TRAIL and DDP of the SGC7901/VCR cells, the IC50 of VCR, DDP, ADM, and 5?Fu treatment was significantly decreased compared with the control group or TRAIL?treated group (P<0.05). After treatment with 0, 10, 50 ng/ml TRAIL in combination with 0. 4 μg/ml DDP, the SGC7901/VCR cells showed significantly higher activation of caspase 3, down?regulation of DNA?PKcs/Akt/GSK?3β signaling pathway, and higher inhibition of MDR1(P?gp) and MRP1 than those treated with TRAIL alone (P<0.01 for all). The mRNA and protein levels of DR4, DR5, c?myc were significantly decreased after silencing c?myc with specific siRNA in the SGC7901/VCR cells ( P<0. 01 for all ) , and were significantly increased after transfection of recombinant plasmid c?myc into the SGC7901/VCR cells (P<0.01 foe all). After the treatment with 10 ng/ml TRAIL, 0. 25 μg/ml DDP+10 ng/ml TRAIL and 0. 5 μg/ml DDP+10 ng/ml TRAIL, the relative expression level of c?myc protein in the SGC7901/VCR cells was 0.314±0.012, 0.735±0.026, and 0.876±0.028, respectively, and the relative expression of cytochrome C was 0.339±0.036, 0.593±0.020 and 0.735±0.031, respectively, and the relative expression levels of DR4, DR5, active?caspase 3 and active?caspase 9 in the SGC7901/VCR cells were also increased along with increasing DDP concentrations. Conclusions The activation of DNA?PKcs/Akt/GSK?3β signaling pathway and high expression of MDR1 and MRP1 play an important role in the multi?drug resistance properties of SGC7901/VCR cells. After combining with TRAIL, DDP can enhance the expression of DR4 and DR5 through up?regulating c?myc and enhancing the activation of caspase 3 and caspase 9 by facilitating mitochondrial release of cytochrome C. It may be an important molecular mechanism of DDP?induced sensitization of TRAIL to reverse the multidrug resistancein SGC7901/VCR cells.

Objective To study the molecular mechanism of cisplatin to enhance the ability of tumor necrosis factor?related apoptosis?inducing ligand ( TRAIL ) in reversing multidrug resistance in vincristine?resistant human gastric cancer SGC7901/VCR cells. Methods MTT assay was used to measure the 50% inhibiting concentration( IC50 ) and cell survival in SGC7901 and SGC7901/VCR cells after different treatments.SGC7901/VCR cells were treated with different concentrations of DDP, different concentrations of TRAIL alone or in combination, and then the mRNA and protein levels of several genes were determined by RT?PCR, RT?qPCR and Western?blot analysis. After targeted silencing with specific siRNA and transfection of recombinant plasmid c?myc into the SGC7901/VCR cells, the mRNA and protein levels of DR4, DR5 and c?myc were determined by RT?PCR and Western?blot analysis. Results After combined treatment with TRAIL and DDP of the SGC7901/VCR cells, the IC50 of VCR, DDP, ADM, and 5?Fu treatment was significantly decreased compared with the control group or TRAIL?treated group (P<0.05). After treatment with 0, 10, 50 ng/ml TRAIL in combination with 0. 4 μg/ml DDP, the SGC7901/VCR cells showed significantly higher activation of caspase 3, down?regulation of DNA?PKcs/Akt/GSK?3β signaling pathway, and higher inhibition of MDR1(P?gp) and MRP1 than those treated with TRAIL alone (P<0.01 for all). The mRNA and protein levels of DR4, DR5, c?myc were significantly decreased after silencing c?myc with specific siRNA in the SGC7901/VCR cells ( P<0. 01 for all ) , and were significantly increased after transfection of recombinant plasmid c?myc into the SGC7901/VCR cells (P<0.01 foe all). After the treatment with 10 ng/ml TRAIL, 0. 25 μg/ml DDP+10 ng/ml TRAIL and 0. 5 μg/ml DDP+10 ng/ml TRAIL, the relative expression level of c?myc protein in the SGC7901/VCR cells was 0.314±0.012, 0.735±0.026, and 0.876±0.028, respectively, and the relative expression of cytochrome C was 0.339±0.036, 0.593±0.020 and 0.735±0.031, respectively, and the relative expression levels of DR4, DR5, active?caspase 3 and active?caspase 9 in the SGC7901/VCR cells were also increased along with increasing DDP concentrations. Conclusions The activation of DNA?PKcs/Akt/GSK?3β signaling pathway and high expression of MDR1 and MRP1 play an important role in the multi?drug resistance properties of SGC7901/VCR cells. After combining with TRAIL, DDP can enhance the expression of DR4 and DR5 through up?regulating c?myc and enhancing the activation of caspase 3 and caspase 9 by facilitating mitochondrial release of cytochrome C. It may be an important molecular mechanism of DDP?induced sensitization of TRAIL to reverse the multidrug resistancein SGC7901/VCR cells.

Objective To evaluate the apparent diffusion coefficient(ADC)values of different bone marrow infiltration patterns in multiple myeloma(MM)patients with MR whole-body diffusion weighted imaging(WB-DWI)and to determine the ADC thresholds for different bone marrow infiltration patterns.Methods Nineteen MM patients diagnosed for the first time were selected.The lesions types of each site(cervical spine,ribs,sternum,humerus,scapula,sacral spine,ilium,femur,thoracic spine,and lumbar spine)after the WB-DWI images were visually evaluated,which were divided into focal group(including focal lesion in combined focal and diffuse infiltration)[region of interest(ROI)=141],pure diffuse infiltration group(ROI=150),diffuse lesion in combined focal and diffuse infiltration group(ROI=127),"salt-and-pepper"group(ROI=54),and normal appearance group(ROI=68).ADC values were measured and compared between each group and the receiver operating characteristic(ROC)curve was drawn to distinguish different patterns of bone marrow infiltration.Results There was no statistically significant difference in ADC values between the diffuse lesion in combined focal and diffuse infiltration group and the"salt-and-pepper"group(P＞0.99),and there was statistically significant difference in ADC values between the other groups(P＜0.05).The ROC curve showed that the area under the curve(AUC)for identifying focal group and the"salt-and-pepper"group was 0.889[95％confidence interval(CI)0.844-0.934],the AUC for identifying pure diffuse infiltration group and the normal appearance group was 0.968(95％CI 0.949-0.987).ADC values were able to accurately and visually differentiate between the different patterns of bone marrow infiltration.Conclusion The ADC values can be used as a quantitative tool to objectively distinguish different bone marrow infiltration patterns in MM patients.

Aim To explore the correlation between carotid atherosclerosis(CAS)and neutrophil/lymphocyte ratio(NLR)in patients with early morning hypertension,and to construct a line chart model to predict the risk of CAS in patients with hypertension in the morning.Methods 255 patients with early morning hypertension hospitalized in the Affiliated Hospital of Chengde Medical College from October 2019 to November 2022 were collected,and their basic data,blood routine and blood biochemical indexes were collected.All selected patients need to improve 24-hour ambulatory blood pressure monitoring and carotid artery color ultrasound detection.According to the presence or absence of CAS,all selected patients were divided into morning hypertension with CAS group(n=197)and morning hypertension without CAS group(n=58).Multivariate Logistic regression analysis was used to explore the risk factors of early morning hypertension with CAS,and to construct and verify an individual line chart model to predict the risk of early morning hypertension pa-tients with CAS.Results The age,NLR,neutrophils(NE),monocytes(MO),white blood cell(WBC),total cho-lesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDLC)increased in the early morning hyperten-sion with CAS group compared with those in the morning hypertension group without CAS,while the HDLC decreased(P＜0.05).The results of multivariate Logistic regression analysis showed that the age,NLR and TC were higher in the early morning hypertension with CAS group than those in the early morning hypertension without CAS group,while HDLC was lower;Age,NLR and TC were independent risk factors of early morning hypertension with CAS,while HDLC was inde-pendent protective factors of morning hypertension with CAS.Based on the results of multivariate Logistic regression anal-ysis,an individualized line chart model for predicting early morning hypertension with CAS was constructed.The area un-der the ROC curve of the line chart model was 0.853(95％CI:0.802-0.904,P＜0.01).The result of Hosmer Leme-show fit test was x2=1.665(P＞0.05).Conclusions There was a positive correlation between NLR and morning hy-pertension with CAS,and NLR was an independent risk factor for morning hypertension with CAS.The individualized line chart model based on age,NLR,TC and HDLC can effectively predict the risk of hypertension with CAS in the early morn-ing,which provides a theoretical basis for early detection and prevention of atherosclerosis.

AIM:To investigate the effect of Bushen formulae(BHF)on bone metabolism and its possible mechanism in ovariectomized rats with high salt intake.METHODS:According to the random number table method,80 SPF-grade Sprague-Dawley rats were divided into sham group,ovariectomy(OVX)group,medium-high-salt diet(MSD)group,high-salt diet(HSD)group,BHF group,BHF with normal saline(BHF+NS)group,BHF+MSD group,and BHF+ HSD group,with 10 rats in each group.After modeling,different diets and BHF formula interventions were administered,and the concentrations of sodium chloride added to MSD group and HSD group were 2%(w/w)and 8%(w/w),respective-ly.The dose of BHF was 7.8 g·kg-1·d-1,once a day,and the treatment lasted for 12 weeks.Bone density,bone microar-chitecture,bone parameters,bone metabolism biomarkers,bone histopathological changes,the expression of epithelial sodium channel α(ENaCα),Na-Cl cotransporter(NCC),and voltage-gated chloride channel 3(ClC-3)proteins in bone tissue were detected in each group.RESULTS:Compared with sham group,the rats in OVX group had reduced bone density and destroyed bone microstructure.Compared with OVX group,the bone microstructure in MSD and HSD groups was more significantly damaged,while the levels of bone formation markers,bone glycoprotein(BGP)and type Ⅰ procolla-gen N-terminal peptide(PINP),were significantly increased in HSD group(P<0.05).Moreover,the levels of bone re-sorption markers,such as amino-terminal cross-linked telopeptides of type Ⅰ collagen(NTX),carboxy-terminal cross-linked telopeptides of type Ⅰ collagen(CTX)and tartrate-resistant acid phosphatase(TRACP),were significantly in-creased(P<0.05),indicating that bone metabolism was in high-conversion state.High-salt diet accelerated the structural destruction of bone trabeculae,and Western blot results showed that high-salt diet caused decreases in the protein expres-sion levels of ENaCα and ClC-3 and an increase in the protein expression level of NCC in femoral tissues(P<0.05).After BHF intervention,the expression of relevant ion channels caused by high salt could be regulated to different degrees.CONCLUSION:Bushen formulae could differentially regulate the expression of relevant ion channels ENaCα,ClC-3,and NCC induced by high salt to different degrees,which has certain ameliorative and therapeutic effects on the imbalance of bone metabolism.

OBJECTIVE: To explore the genetic etiology of recurrent hydatidiform mole (RHM) and provide accurate guidance for reproduction. METHODS: Peripheral venous blood samples of the probands with RHM and members from 5 unrelated pedigrees were collected. Genomic DNA was extracted by using routine method, and whole exome sequencing was carried out to detect variants of RHM-associated genes including NLRP7 and KHDC3L. Sanger sequencing and real-time quantitative PCR (RT-qPCR) were used to validate the candidate variants and delineate their parental origin. RESULTS: Homozygous or compound heterozygous variants of the NLRP7 gene were identified in four patients from three pedigrees, which included a homozygous deletion of exon 1 to 4 of NLRP7 in patient P1 and her elder sister, compound heterozygous variants of NLRP7 c.939delG (p.Q314Sfs*6) pat and c.1533delG (p.N512Tfs*4) mat in patient P2, and compound heterozygous variants of NLRP7 c.2389_2390delTC (p.A798Qfs*6) pat and c.2165A>G (p.D722G) mat in patient P4. All variants were interpreted as pathogenic or likely pathogenic according to the American College of Medical and Genomics (ACMG) guidelines. Among these, NLRP7 exons 1 to 4 deletion, c.939delG (p.Q314Sfs*6), c.1533delG (p.N512Tfs*4) and c.2389_2390delTC (p.A798Qfs*6) were unreported previously. CONCLUSION Variants of the NLRP7 gene probably underlay autosomal recessive RHM in the three pedigrees, and definitive molecular diagnosis is beneficial for accurate genetic counseling. Above finding has also enriched the spectrum of the NLRP7 variants underlying RHM.
Adaptor Proteins, Signal Transducing/genetics* ; Aged ; China ; Female ; Homozygote ; Humans ; Hydatidiform Mole/pathology* ; Mutation ; Pedigree ; Pregnancy ; Sequence Deletion

OBJECTIVETo study the molecular mechanism of cisplatin to enhance the ability of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in reversing multidrug resistance in vincristine-resistant human gastric cancer SGC7901/VCR cells.

METHODSMTT assay was used to measure the 50% inhibiting concentration (IC₅₀) and cell survival in SGC7901 and SGC7901/VCR cells after different treatments. SGC7901/VCR cells were treated with different concentrations of DDP, different concentrations of TRAIL alone or in combination, and then the mRNA and protein levels of several genes were determined by RT-PCR, RT-qPCR and Western-blot analysis. After targeted silencing with specific siRNA and transfection of recombinant plasmid c-myc into the SGC7901/VCR cells, the mRNA and protein levels of DR4, DR5 and c-myc were determined by RT-PCR and Western-blot analysis.

RESULTSAfter combined treatment with TRAIL and DDP of the SGC7901/VCR cells, the IC₅₀ of VCR, DDP, ADM, and 5-Fu treatment was significantly decreased compared with the control group or TRAIL-treated group (P < 0.05). After treatment with 0, 10, 50 ng/ml TRAIL in combination with 0.4 µg/ml DDP, the SGC7901/VCR cells showed significantly higher activation of caspase 3, down-regulation of DNA-PKcs/Akt/GSK-3β signaling pathway, and higher inhibition of MDR1(P-gp) and MRP1 than those treated with TRAIL alone (P < 0.01 for all). The mRNA and protein levels of DR4, DR5, c-myc were significantly decreased after silencing c-myc with specific siRNA in the SGC7901/VCR cells (P < 0.01 for all), and were significantly increased after transfection of recombinant plasmid c-myc into the SGC7901/VCR cells (P < 0.01 foe all). After the treatment with 10 ng/ml TRAIL, 0.25 µg/ml DDP + 10 ng/ml TRAIL and 0.5 µg/ml DDP + 10 ng/ml TRAIL, the relative expression level of c-myc protein in the SGC7901/VCR cells was 0.314 ± 0.012, 0.735 ± 0.026, and 0.876 ± 0.028, respectively, and the relative expression of cytochrome C was 0.339 ± 0.036, 0.593 ± 0.020 and 0.735 ± 0.031, respectively, and the relative expression levels of DR4, DR5, active-caspase 3 and active-caspase 9 in the SGC7901/VCR cells were also increased along with increasing DDP concentrations.

CONCLUSIONSThe activation of DNA-PKcs/Akt/GSK-3β signaling pathway and high expression of MDR1 and MRP1 play an important role in the multi-drug resistance properties of SGC7901/VCR cells. After combining with TRAIL, DDP can enhance the expression of DR4 and DR5 through up-regulating c-myc and enhancing the activation of caspase 3 and caspase 9 by facilitating mitochondrial release of cytochrome C. It may be an important molecular mechanism of DDP-induced sensitization of TRAIL to reverse the multidrug resistancein SGC7901/VCR cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; pharmacology ; Down-Regulation ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Fluorouracil ; administration & dosage ; pharmacology ; Formazans ; Genes, myc ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Inhibitory Concentration 50 ; Multidrug Resistance-Associated Proteins ; metabolism ; Neoplasm Proteins ; metabolism ; Plasmids ; Proto-Oncogene Proteins c-myc ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Stomach Neoplasms ; drug therapy ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; administration & dosage ; pharmacology ; Tetrazolium Salts ; Transfection ; methods