Background and purpose:Polo-like kinase 4(PLK4)is a cell cycle regulatory protein,which is closely related to the occurrence and development of a variety of malignant tumors.The aim of this study was to investigate the role of PLK4 in the proliferation,invasion and apoptosis of oral squamous cell carcinoma(OSCC).Methods:Gene Expression Profiling Interactive Analysis 2(GEPIA2)and the University of Alabama at Birmingham Cancer Data Analysis Portal(UALCAN)were used to analyze the expression of PLK4 in head and neck squamous cell carcinoma and surrounding normal tissues.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot were employed to evaluate the mRNA and protein expression of PLK4 in OSCC cells.We further analyzed PLK4,phosphoinositide 3-kinase(PI3K),phosphorylated phosphoinositide 3-kinase(p-PI3K),protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),survivin,and cyclin D1 protein expression.The effects of PLK4 on cell proliferation,apoptosis and invasion were analyzed by cell counting kit-8(CCK-8)assay,flow cytometry and transwell assay.In addition,12 4-week-old SPF BALB/c female nude mice were divided into sh-NC group(n=6)and sh-PLK4 group(n=6)by random number table method.The sh-NC/sh-PLK4 cells were injected into the right anterior armpit of nude mice for subcutaneous tumor formation.The body weight,tumor volume and tumor weight of the two groups of nude mice were observed,and the stripped tumor tissues were analyzed by H-E staining.The animal experiments were approved by the Animal Experiment Center of Shandong Second Medical University(No:2024SDL840).Results:The results of GEPIA2 and UALCAN online databases showed that PLK4 was highly expressed in OSCC compared with normal tissues.In addition,PLK4 was highly expressed in OSCC cell lines(HN6,Cal-27,SCC-4,SCC-9,SCC-25)compared with oral mucosal epithelial cells(P＜0.05).Western blot results showed that the expression levels of PI3K/AKT signaling pathway proteins p-PI3K,p-AKT,cyclin D1 and survivin were decreased after PLK4 knockdown and increased after PLK4 overexpression in OSCC cells(P＜0.05).The cell proliferation activity and the number of transmembrane cells were positively correlated with the decrease and increase of PLK4 expression(P＜0.05),while the cell apoptotic rate was negatively correlated(P＜0.05),indicating that cell proliferation and apoptosis were both affected.In addition,compared with the sh-NC group,the tumor volume and weight of the nude mice in the sh-PLK4 group were decreased(P＜0.05),while there was no significant difference in the body weight of the nude mice between the two groups(P＞0.05).Moreover,the nuclear atypia of tumor tissues in sh-PLK4 group was lower than that in sh-NC group(P＜0.05).Conclusion:PLK4 can regulate the invasion,proliferation and apoptosis of OSCC cells,potentially through the activation of PI3K/AKT signaling pathway.

Background and purpose:Polo-like kinase 4(PLK4)is a cell cycle regulatory protein,which is closely related to the occurrence and development of a variety of malignant tumors.The aim of this study was to investigate the role of PLK4 in the proliferation,invasion and apoptosis of oral squamous cell carcinoma(OSCC).Methods:Gene Expression Profiling Interactive Analysis 2(GEPIA2)and the University of Alabama at Birmingham Cancer Data Analysis Portal(UALCAN)were used to analyze the expression of PLK4 in head and neck squamous cell carcinoma and surrounding normal tissues.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot were employed to evaluate the mRNA and protein expression of PLK4 in OSCC cells.We further analyzed PLK4,phosphoinositide 3-kinase(PI3K),phosphorylated phosphoinositide 3-kinase(p-PI3K),protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),survivin,and cyclin D1 protein expression.The effects of PLK4 on cell proliferation,apoptosis and invasion were analyzed by cell counting kit-8(CCK-8)assay,flow cytometry and transwell assay.In addition,12 4-week-old SPF BALB/c female nude mice were divided into sh-NC group(n=6)and sh-PLK4 group(n=6)by random number table method.The sh-NC/sh-PLK4 cells were injected into the right anterior armpit of nude mice for subcutaneous tumor formation.The body weight,tumor volume and tumor weight of the two groups of nude mice were observed,and the stripped tumor tissues were analyzed by H-E staining.The animal experiments were approved by the Animal Experiment Center of Shandong Second Medical University(No:2024SDL840).Results:The results of GEPIA2 and UALCAN online databases showed that PLK4 was highly expressed in OSCC compared with normal tissues.In addition,PLK4 was highly expressed in OSCC cell lines(HN6,Cal-27,SCC-4,SCC-9,SCC-25)compared with oral mucosal epithelial cells(P＜0.05).Western blot results showed that the expression levels of PI3K/AKT signaling pathway proteins p-PI3K,p-AKT,cyclin D1 and survivin were decreased after PLK4 knockdown and increased after PLK4 overexpression in OSCC cells(P＜0.05).The cell proliferation activity and the number of transmembrane cells were positively correlated with the decrease and increase of PLK4 expression(P＜0.05),while the cell apoptotic rate was negatively correlated(P＜0.05),indicating that cell proliferation and apoptosis were both affected.In addition,compared with the sh-NC group,the tumor volume and weight of the nude mice in the sh-PLK4 group were decreased(P＜0.05),while there was no significant difference in the body weight of the nude mice between the two groups(P＞0.05).Moreover,the nuclear atypia of tumor tissues in sh-PLK4 group was lower than that in sh-NC group(P＜0.05).Conclusion:PLK4 can regulate the invasion,proliferation and apoptosis of OSCC cells,potentially through the activation of PI3K/AKT signaling pathway.

OBJECTIVE: To assess the efficacy of Qingda Granule (QDG) in ameliorating hypertension-induced cardiac damage and investigate the underlying mechanisms involved. METHODS: Twenty spontaneously hypertensive rats (SHRs) were used to develope a hypertension-induced cardiac damage model. Another 10 Wistar Kyoto (WKY) rats were used as normotension group. Rats were administrated intragastrically QDG [0.9 g/(kg•d)] or an equivalent volume of pure water for 8 weeks. Blood pressure, histopathological changes, cardiac function, levels of oxidative stress and inflammatory response markers were measured. Furthermore, to gain insights into the potential mechanisms underlying the protective effects of QDG against hypertension-induced cardiac injury, a network pharmacology study was conducted. Predicted results were validated by Western blot, radioimmunoassay immunohistochemistry and quantitative polymerase chain reaction, respectively. RESULTS: The administration of QDG resulted in a significant decrease in blood pressure levels in SHRs (P<0.01). Histological examinations, including hematoxylin-eosin staining and Masson trichrome staining revealed that QDG effectively attenuated hypertension-induced cardiac damage. Furthermore, echocardiography demonstrated that QDG improved hypertension-associated cardiac dysfunction. Enzyme-linked immunosorbent assay and colorimetric method indicated that QDG significantly reduced oxidative stress and inflammatory response levels in both myocardial tissue and serum (P<0.01). CONCLUSIONS Both network pharmacology and experimental investigations confirmed that QDG exerted its beneficial effects in decreasing hypertension-induced cardiac damage by regulating the angiotensin converting enzyme (ACE)/angiotensin II (Ang II)/Ang II receptor type 1 axis and ACE/Ang II/Ang II receptor type 2 axis.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Hypertension/pathology* ; Renin-Angiotensin System/drug effects* ; Rats, Inbred SHR ; Oxidative Stress/drug effects* ; Male ; Rats, Inbred WKY ; Blood Pressure/drug effects* ; Myocardium/pathology* ; Rats ; Inflammation/pathology*

A new multifunctional fluorescent"turn-on"biosensor was constructed based on a guanine-rich hairpin probe and G-quadruplex/hemin DNAzyme.The probe was labeled with quencher and fluorescein at 5′and 3′terminus,respectively.In the absence of the target,the probe form hairpin structure and the fluorescence of the fluorescein was quenched by the quencher.However,in the presence of the target,the hairpin structure transformed to G-quadruplex and the fluorescence was recovered.By monitoring the fluorescence of the probe before and after combining with the target,both hemin and hemoglobin were detected rapidly and conveniently with high sensitivity.The limit of detection for hemin and hemoglobin were estimated to be 283 pmol/L and 109 pmol/L,respectively.The specific interaction between the DNA probe and hemin were investigated via absorption and fluorescence spectra.Furthermore,L-cysteine was detected with naked eyes based on the peroxidase-like activity of the new G-quadruplex/hemin DNAzyme.The limit of detection for L-cysteine was estimated to be 3.6 μmol/L,which was lower than the normal level of intracellular L-cysteine(30-200 μmol/L).The method developed here was rapid,simple and sensitive for detection of hemin,hemoglobin and L-cysteine,demonstrating great potential in bioanalysis,drug screening,and disease diagnosis.

Objective To analyze the expression of N-MYC and N-MYC downstream regulated gene-1(NDRG1)in gastric cancer tissues,and to assess their effects on biological characteristics of gastric cancer cells.Methods Paired of gastric cancer tissues and adjacent normal tissues resected from 82 cases of patholog-ically confirmed gastric cancer who underwent surgical treatment in the hospital from January 2021 to May 2023 were collected.Gastric cancer tissues and adjacent normal tissues of 82 patients who were surgically re-sected and pathologically diagnosed with gastric cancer in the hospital from January 2021 to May 2023 were collected.Real-time quantitative PCR(qPCR)was used to detect the relative mRNA expression levels of N-MYC and NDRG1,and clinical data of the patients were collected.The correlation between the mRNA expres-sion of N-MYC and NDRG1 and clinicopathological features of the patients was discussed.NCI-N87 cells in logarithmic growth phase were selected and cultured in vitro.N-MYC interference plasmid(si-N-MYC)and its negative control(si-NC)was transfected into NCI-N87 cells,respectively,which were recorded as si-NC group and si-N-MYC group.Moreover,si-N-MYC was co-transfected into NCI-N87 cells with anti-NC and an-ti-NDRG1,respectively,and denoted as si-N-MYC+anti-NC group and si-N-MYC+anti-NDRG1 group.CCK-8 assay was used to detect cell proliferation activity,Transwell assay was used to detect cell invasion ability,and Western blotting assay was used to detect N-MYC and NDRG1 protein expression in cells.Results The relative expression of N-MYC mRNA in gastric cancer tissues was higher than that in paracancer tissues(P＜0.05),and the relative expression of NDRG1 mRNA was lower than that in paracancer tissues(P＜0.05).There were significant differences in the expression of N-MYC and NDRG1 mRNA in patients with different TNM stages,lymph node metastasis and distant metastasis(P＜0.05).Compared with the si-NC group,the cell proliferation and invasion ability of the si-N-MYC group were decreased(P＜0.05),and the expression of NDRG1 protein was down-regulated(P＜0.05).Compared with si-N-MYC+anti-NC group,cell proliferation and invasion ability of si-N-MYC+anti-NDRG1 group were increased(P＜0.05).N-MYC could target and regulate NDRG1,and knocking down NDRG1 could reverse the biological effects of N-MYC on gastric cells.Conclusion In gastric cancer tissue,N-MYC mRNA expression is upregulated and NDRG1 mRNA expression is downregulated,both of which play important roles in the regulation of malignant biological behaviors such as proliferation and invasion of gastric cancer cells.

Since Curcumae Radix decoction pieces have multiple sources, it is difficult to distinguish depending on traditional cha-racters, and the mixed use of multi-source Curcumae Radix will affect its clinical efficacy. Heracles Neo ultra-fast gas phase electronic nose was used in this study to quickly identify and analyze the odor components of 40 batches of Curcumae Radix samples from Sichuan, Zhejiang, and Guangxi. Based on the odor fingerprints established for Curcumae Radix decoction pieces of multiple sources, the odor components was identified and analyzed, and the chromatographic peaks were processed and analyzed to establish a rapid identification method. Principal component analysis(PCA), discriminant factor analysis(DFA), and soft independent modeling cluster analysis(SIMCA) were constructed for verification. At the same time, one-way analysis of variance(ANOVA) combined with variable importance in projection(VIP) was employed to screen out the odor components with P<0.05 and VIP>1, and 13 odor components such as β-caryophyllene and limonene were hypothesized as the odor differential markers of Curcumae Radix decoction pieces of diffe-rent sources. The results showed that Heracles Neo ultra-fast gas phase electronic nose can well analyze the odor characteristics and rapidly and accurately discriminate Curcumae Radix decoction pieces of different sources. It can be applied to the quality control(e.g., online detection) in the production of Curcumae Radix decoction pieces. This study provides a new method and idea for the rapid identification and quality control of Curcumae Radix decoction pieces.
Drugs, Chinese Herbal/analysis* ; Electronic Nose ; China ; Plant Roots/chemistry* ; Limonene/analysis* ; Chromatography, High Pressure Liquid

Objective: To evaluate the long-term outcomes, failure patterns and prognostic factors of definitive radiotherapy in patients with cervical esophageal carcinoma (CEC). Methods: We retrospectively reviewed the clinical data of 148 CEC patients who treated with definitive radiotherapy in Cancer Hospital of Chinese Academy of Medical Sciences from January 2001 to December 2017. The median radiation dose was 66 Gy (59.4-70 Gy) and 33.1% of patients received concurrent chemotherapy. The Kaplan-Meier method was used to calculate survival rates. The log rank test was used for survival comparison and univariate prognostic analysis. The Cox model was used for multivariate prognostic analysis. Results: The median follow-up time was 102.6 months. The median survival time, 2- and 5-year overall survival (OS) were 22.7 months, 49.9% and 28.3%. The median, 2- and 5-year progression-free survival were 12.6 months, 35.8% and 25.8%. The 2- and 5-year locoregional recurrence-free survival were 59.1% and 50.8%. The 2- and 5-year distant metastases-free survival were 74.6% and 65.9%. Multivariate analysis showed that EQD(2)>66 Gy was the only independent prognostic indicator for OS (P=0.040). The median survival time and 5-year OS rate significantly improved in patients who received EQD(2)>66 Gy than those who received≤66 Gy (31.2 months vs. 19.2 months, 40.1% vs. 19.1%, P=0.027). A total of 87 patients (58.8%) developed tumor progression. There were 50 (33.8%), 23 (15.5%) and 39 (26.4%) patients developed local, regional recurrence and distant metastases, respectively. Eleven patients (7.4%) underwent salvage surgery, and the laryngeal preservation rate for entire group was 93.9%. Conclusions: Definitive radiotherapy is an effective treatment for cervical esophageal carcinoma with the advantage of larynx preservation. Local recurrence is the major failure pattern. EQD(2)>66 Gy is associated with the improved overall survival.
Humans ; Retrospective Studies ; Esophageal Neoplasms/pathology* ; Carcinoma/drug therapy* ; Prognosis ; Treatment Outcome ; Chemoradiotherapy/methods* ; Radiotherapy Dosage

Objective:To explore the possible mechanism of electroacupuncture to improve detrusor hyperreflex after suprasacral spinal cord injury. Methods:A total of 60 female Sprague-Dawley rats were included. According to the random number table, twelve were selected as the blank group, twelve as the sham operation group, and the remaining 36 were made neurogenic bladder models using modified T₁₀ spinal cord transection. After that, twelve of them were randomly selected as the model group and twelve were as the electroacupuncture group from the model rats that met the requirements. On the 19th day after modelling, Ciliao (BL32), Zhongji (RN3) and Sanyinjiao (SP6) were taken for electroacupuncture. After seven days of continuous treatment, urodynamic testing was performed, content of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in detrusor was determined by ELISA, and the level of phosphorylation of myosin light chain kinase (p-MLCK) of detrusor was determined by Western blotting. Results:Compared with the blank group and the sham operation group, the maximum bladder capacity and bladder compliance significantly reduced (P < 0.01), and the base pressure and leakage point pressure of bladder significantly increased (P < 0.01); the content of cAMP and PKA in detrusor reduced (P < 0.01), p-MLCK in detrusor reduced (P < 0.05) in the model group. Compared with the model group, the maximum bladder capacity and bladder compliance increased (P < 0.01), the base pressure of the bladder and the pressure at the leak point decreased (P < 0.05); the contents of cAMP and PKA protein in detrusor increased (P < 0.05), the p-MLCK in detrusor increased (P < 0.05) in the electroacupuncture group. Conclusion:Electroacupuncture at Ciliao, Zhongji and Sanyinjiao points could improve the bladder function of rats with detrusor hyperreflex after complete spinal cord injury, and its mechanism may be related to up-regulating the expression of cAMP and PKA, phosphorylating and inactivating p-MLCK, which promote relaxation of detrusor.

Objective To investigate the relationship between polymorphisms of gene promoter region INS 5′UTR single nu-cleotide and type 2 diabetes and serum IAA-Ab levels.Methods By Sequenom MassArray SNP genotyping detection technology, INS 3 pyomter regime single nucleotide polymorphisms (rs689,rs714641 77 and rs3842738)of 497 patients in Chongqing with type 2 diabetes cases(treatment group)and 500 cases(control group)were genotyped and analyzed.IAA-Ab levels in diabetes patients was detected.Theχ2 test statistic was used to analyze the treatment group and control groups.The genotype frequency distribution of IAA-Ab-positive and negative groups SNP was analyzed by non-conditional logistic regression,adjusted for sex,age impact,cal-culated the odds ratio (OR)and 95 % confidence interval(CI ).The polymorphic loci with type 2 diabetes susceptibility and serum GAD-Ab levels was evaluated.Results The genotype frequency distribution of rs689AA,TT and AT was 58.75%,28.77% and 12.47%,respectively.The control group are 50.40%,35.60% and 14.00% respectively.The difference was statistically significant (χ2 =3.923,P <0.05).Compared with the genotype of AA,TT genotype can decrease risky of diabetes,with OR values 0.35(95%CI :0.18-1.06).There was significant difference of AA,TT,AT genotypes between IAA-Ab negative and IAA-Ab positive pa-tients (P < 0.05 ).Conclusion INS polymorphisms might be related to the risky of type 2 diabetes and serum IAA-Ab level in chinses population.

Objective To study the variants of S gene of hepatitis B virus among the breakthrough infection children immunized with hepatitis B vaccine in Huzhou city.Methods HBV DNA S gene regions were amplified and sequenced in 29 children with HBsAg-positive/HBV DNA-positive after immunized with hepatitis B vaccine.Results The sequencing results showed that 26 cases were B genotypes,3 were C genotypes.The amino acids mutations were presented in 9 children serum HBV DNA.The major amino acid mutations were in the "a" determinant of HBV DNA S gene.Conclusion Different S gene variants maybe cause HBV vaccination failure in some children in Huzhou city.No predominant variant was found.