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.

BACKGROUND:Studies have found that massage can reduce blood sugar,promote myogenic factor expression,and increase skeletal muscle content.The extracellular matrix is an important component of skeletal muscle,and association between massage and extracellular matrix and their mechanism of action are still unclear.OBJECTIVE:To explore the effect of massage on extracellular matrix collagen deposition in type 2 diabetic sarcopenia rats.METHODS:Totally 24 Wistar male rats were randomly divided into blank group,model group,and massage group.High-fat diet combined with the streptozotocin method was used to establish a type 2 diabetes mellitus and sarcopenia model.After successful model establishment,the massage group used abdominal massage combined with hind limbs.After 8 weeks of treatment,the fasting blood glucose and serum insulin levels of the rats were measured.The skeletal muscle mass was detected by dual-energy X-ray.The exhaustion time was measured by small animal treadmill.The sliding angle was measured by inclined board test.The pathological changes of skeletal muscle tissue were observed by hematoxylin-eosin staining.The skeletal muscle collagen deposition was observed by Masson staining.The mRNA and protein expressions of type Ⅰ and type Ⅲ collagen in skeletal muscle were detected by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)Compared with the model group,the blood glucose(P＜0.05)and serum insulin(P＜0.01)decreased in the massage group.(2)Compared with the model group,the skeletal muscle mass,running exhaustion time,and the angle of inclined plate experiment were increased in massage group(P＜0.05).(3)Compared with the model group,the skeletal muscles of the massage group were arranged neatly,muscle atrophy was improved,and collagen fiber deposition was reduced.(4)Compared with the model group,the expression levels of type Ⅰ and type Ⅲ collagen mRNA and protein in skeletal muscle were decreased in the massage group(P＜0.05).(5)The results suggest that massage can enhance insulin sensitivity,lower blood sugar,improve skeletal muscle mass,strength and function,and diminish collagen deposition in rats with type 2 diabetes,and may be a potential target for massage to exert its therapeutic effects.

OBJECTIVES: This study aimed to explore the therapeutic effect and mechanism of ginsenoside Rb3 on experimental periodontitis and bone resorption in rats. METHODS: Male SD rats were randomly divided into a control group, a ligation group, an Rb3 group, and a doxycycline (Dox) group for in vivo experiments. A periodontitis model was established by ligating the maxillary second molar, and samples were collected after 3 weeks of drug treatment. Micro-CT assessment of alveolar bone resorption was performed, and hematoxylin-eosin (HE) staining was used to observe pathological changes in periodontal and visceral tissues. Tartrate resistant acid phosphatase (TRAP) staining was applied to detect the formation of osteoclasts in periodontal tissues, and enzyme-linked immunosorbent assay (ELISA) was adopted to detect the serum levels of interleukin (IL)-6, IL-8, immunoglobulin (Ig)M, and IgG. Quantitative polymerase chain reaction (qPCR) was employed to detect the expression of factors related to gingival inflammation and osteoclast formation. Immunofluorescence staining was used to detect phospho-extracellular signal-regulated kinase (p-ERK) expression. In vitro experiments were conducted by pretreating RAW264.7 cells with drugs and adding lipopolysaccharides (LPS) stimulation from Porphyromonas gingivalis (P. gingivalis). IL-1β and IL-6 mRNA expression was detected by qPCR, and Western blot was used to detect the effect of Rb3 on the mitogen-activated protein kinases (MAPKs) signaling pathway. RESULTS: Compared with the control group, the ligation group showed significant periodontitis and bone resorption. Compared with the ligation group, the Rb3 group showed a decrease in alveolar bone resorption and osteoclast formation; p-ERK/ERK ratio, IL-1β, IL-6, and nuclear factor of activated T cells (NFATc1) mRNA levels and downstream gene expression in periodontal tissues; serum IL-6, IL-8, IgG, and IgM levels. Rb3 reduced IL-8 and IL-1β mRNA expression levels and p-ERK/ERK and p-p38 MAPK/p38 MAPK ratios in RAW264.7 cells induced by P. gingivalis LPS stimulation. CONCLUSIONS Rb3 inhibits inflammation and bone resorption in experimental periodontitis in rats. Compared with Dox, Rb3 has better effects in inhibiting pro-inflammatory factors and osteoclast gene expression and may exert anti-inflammatory effects by activating the MAPK signaling pathway.
Animals ; Ginsenosides/therapeutic use* ; Rats, Sprague-Dawley ; Male ; Periodontitis/pathology* ; Rats ; Osteoclasts/drug effects* ; Interleukin-1beta/metabolism* ; Interleukin-6/blood* ; Mice ; Alveolar Bone Loss ; Interleukin-8/blood* ; Immunoglobulin G/blood* ; RAW 264.7 Cells ; Transcription Factors

BACKGROUND:Studies have found that massage can reduce blood sugar,promote myogenic factor expression,and increase skeletal muscle content.The extracellular matrix is an important component of skeletal muscle,and association between massage and extracellular matrix and their mechanism of action are still unclear.OBJECTIVE:To explore the effect of massage on extracellular matrix collagen deposition in type 2 diabetic sarcopenia rats.METHODS:Totally 24 Wistar male rats were randomly divided into blank group,model group,and massage group.High-fat diet combined with the streptozotocin method was used to establish a type 2 diabetes mellitus and sarcopenia model.After successful model establishment,the massage group used abdominal massage combined with hind limbs.After 8 weeks of treatment,the fasting blood glucose and serum insulin levels of the rats were measured.The skeletal muscle mass was detected by dual-energy X-ray.The exhaustion time was measured by small animal treadmill.The sliding angle was measured by inclined board test.The pathological changes of skeletal muscle tissue were observed by hematoxylin-eosin staining.The skeletal muscle collagen deposition was observed by Masson staining.The mRNA and protein expressions of type Ⅰ and type Ⅲ collagen in skeletal muscle were detected by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)Compared with the model group,the blood glucose(P＜0.05)and serum insulin(P＜0.01)decreased in the massage group.(2)Compared with the model group,the skeletal muscle mass,running exhaustion time,and the angle of inclined plate experiment were increased in massage group(P＜0.05).(3)Compared with the model group,the skeletal muscles of the massage group were arranged neatly,muscle atrophy was improved,and collagen fiber deposition was reduced.(4)Compared with the model group,the expression levels of type Ⅰ and type Ⅲ collagen mRNA and protein in skeletal muscle were decreased in the massage group(P＜0.05).(5)The results suggest that massage can enhance insulin sensitivity,lower blood sugar,improve skeletal muscle mass,strength and function,and diminish collagen deposition in rats with type 2 diabetes,and may be a potential target for massage to exert its therapeutic effects.

OBJECTIVE To investigate the effects of medicated serum of Siwutang on autophagy of ovarian granulosa cells(KGN cells)in polycystic ovarian syndrome(PCOS)and its underlying mechanism.METHODS Blank serum and different-concentration medicated serum of Siwutang were prepared by intragastric administration of normal saline and different doses of Siwutang[0.52,1.04,2.08 g/(kg·d)]in 3-month-old female SD rats.After screening the intervention concentration of Siwutang medicated serum,KGN cells were divided into control group(without any treatment),dehydroepiandrosterone(DHEA)group(treated with 50 μmol/L DHEA for 48 h),blank serum group(treated with 50 μmol/L DHEA for 48 h and with 10％blank serum for 72 h)and medium-concentration of Siwutang medicated serum group(treated with 50 μmol/L DHEA for 48 h and with 10％medium-concentration Siwutang medicated serum for 72 h).The number of autophagosomes was observed in each group,and protein expressions of pathway-related proteins[fructose-1,6-bisphosphatase 1(FBP1),mammalian target of rapamycin(mTOR),phosphorylated mTOR(p-mTOR)],autophagy-related proteins[p62,microtubule-associated protein 1 light chain 3(LC3)]and mRNA expression of FBP1 were also detected.The(transfected)cells were further divided into Siwutang group(treated with 10％medium dose of Siwutang medicated serum for 72 h after 48 h intervention with 50 μmol/L DHEA),Siwutang+si-NC group[negative control small interfering RNA(siRNA)transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration of Siwutang medicated serum for 72 h]and Siwutang+si-FBP1 group(FBP1 siRNA transfected cells treated with 50 μmol/L DHEA for 48 h,and then with 10％medium-concentration Siwutang medicated serum for 72 h).The effects of knocking down FBP1 on the above-mentioned effects of Siwutang were detected.RESULTS Compared with control group,DHEA group exhibited an increase in the number of autophagosomes,an elevated LC3-Ⅱ/LC3-Ⅰ and p-mTOR/mTOR,as well as increases in protein and mRNA expressions of FBP1,and decreased protein expression of p62(P＜0.05).Compared to both DHEA group and blank serum group,the medium-concentration of Siwutang medicated serum group showed a decrease in the number of autophagosomes,a decrease in LC3-Ⅱ/LC3-Ⅰ,and increases in p-mTOR/mTOR,protein expression of p62,protein and mRNA expressions of FBP1(P＜0.05).After knocking down FBP1,compared with Siwutang+si-NC group,Siwutang+si-FBP1 group showed a significant decrease in cell viability,protein expression of p62,protein and mRNA expressions of FBP1 as well as p-mTOR/mTOR,and an increase in LC3-Ⅱ/LC3-Ⅰ(P＜0.05).CONCLUSIONS Siwutang can promote the phosphorylation of mTOR protein by up-regulating the protein and mRNA expressions of FBP1 in KGN cells,thus inhibiting autophagy of KGN cells.

Tuberous sclerosis complex(TSC)is a rare genetic disease that can lead to benign dysplasia in multiple organs such as the skin, brain, eyes, oral cavity, heart, lungs, kidneys, liver, and bones. Its main symptoms include epilepsy, intellectual disabilities, skin depigmentation, and facial angiofibromas, whilst incidence is approximately 1 in 10 000 to 1 in 6000 newborns. This case presents a middle-aged woman who initially manifested with epilepsy and nodular depigmentation. Later, she developed a lower abdominal mass, elevated creatinine, and severe anemia. Based on clinical features and whole exome sequencing, the primary diagnosis was confirmed as TSC. Laboratory and imaging examinations revealed that the lower abdominal mass originated from the uterus. CT-guided biopsy pathology and surgical pathology suggested a combination of leiomyoma and abscess. With the involvement of multiple organs and various complications beyond the main diagnosis, the diagnostic and therapeutic process for this patient highlights the importance of rigorous clinical thinking and multidisciplinary collaboration in the diagnosis and treatment of rare and challenging diseases.

Objective : To investigate the effects of luteolin on invasion and migration of endometriosis stromal cells and whether its mechanism is related to the regulation of 15-hydroxyprostaglandin dehydrogenase (HPGD) expres- sion． Methods : The endometrial stromal cells HEM15A were divided into control group (cells were cultured in nor- mal medium) ，luteolin group ( cells were treated with different concentrations of luteolin) ，si-HPGD group ( cells were transfected with si-HPGD) ，si-NC group ( cells were transfected with si-NC ) ，luteolin + si-HPGD Group (cells were transfected with si-HPGD and treated with 20 μmol / L luteolin) ，Luteolin + si-NC Group ( cells were transfected with si-NC and treated with 20 μmol / L luteolin) . Real-time quantitative PCR was used to detect mRNA level of HPGD．Cell proliferation was detected by CCK-8 assay，Transwell and scratch assays were used to detect cell invasion and migration．The protein expression of proliferating cell nuclear antigen (PCNA) ，matrix metallopro- teinase (MMP-2) ，MMP-9 and HPGD were detected by Western blot，and the level of prostaglandin E2 ( PGE2) was detected by ELISA. Results : Compared with 0 μmol / L luteolin，luteolin at 20，50 and 100 μmol / L significantly inhibited the proliferation activity of hEM15A cells，and reduced PCNA expression ( all P＜0. 05) ．Compared with the control group，20 μmol / L of luteolin significantly inhibited cell invasion and migration (P ＜0. 05) ，decreased the expressions of MMP-2 and MMP-9 (P＜0. 05) ，and up-regulated the mRNA and protein levels of HPGD (P < 0. 05) ，while inhibited cellular PGE2 level (P＜0. 05) ．Compared with the luteolin group，the luteolin + si-HPGD group increased cell invasion and migration (P ＜0. 05 ) ，increased the expressions of MMP-2 and MMP-9 (P < 0. 05) ． Conclusion Luteolin regulates HPGD / PGE2 signaling pathway to inhibit the invasion and migration of en- dometrial stromal cells.

OBJECTIVE To evaluate the quality of Prunella vulgaris by entropy weight method combined with grey correlation degree analysis method by determining the contents of rosmarinic acid， total flavonoids， water and water-soluble extract. METHODS The content of rosmarinic acid was determined by HPLC； the content of total flavonoids was determined by UV spectrophotometry； the contents of water and water-soluble extract were determined according to the general rules of Chinese Pharmacopoeia （part Ⅳ）. The weight coefficients of four indexes as rosmarinic acid were calculated by entropy weight method， and the relative correlation degree of each index was analyzed by grey correlation degree analysis method， and the quality of 117 batches of P. vulgaris was ranked. RESULTS The contents of rosmarinic acid， total flavonoids， water and water-soluble extractive were 0.237 1%-0.943 0%， 2.923 6%-6.400 7%， 8.736 7%-12.446 3%， 11.638 8%-19.166 2%， respectively. The weight coefficients were 0.297 1， 0.277 5， 0.163 7 and 0.261 6， respectively. The relative correlation degrees were 0.159 3-0.616 3， and the correlation degree of S24 batch of samples was the highest （0.616 3）； that of P. vulgaris from Nanjing of Jiangsu Province ranged from 0.500 9 to 0.616 3， ranking the first. CONCLUSIONS Using the contents of rosmarinic acid， total flavonoids， water and water-soluble extract as indexes， entropy weight method and grey correlation degree analysis method can be used to evaluate the quality of P. vulgaris. The quality of different batches of P. vulgaris are different， and the quality of P. vulgaris produced in Nanjing of Jiangsu Province is the best.

Objective:To evaluate the effects of comprehensive rehabilitation nursing based on guided education and training on rehabilitation treatment in school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection.Methods:Fifty movement disorders school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection in Institute of Functional Neurosurgery, Xuanwu Hospital, Capital Medical University from January 2017 to December 2019 were enrolled in this study. The child patients were divided into the control group and the observation group according to the order of admission, with 25 cases in each group, and the control group was from January 2017 to June 2018 while the observation group was from July 2018 to December 2019. In the control group, routine nursing after epileptogenic focus resection and functional exercise were conducted. The observation group was treated with comprehensive rehabilitation nursing based on guided education and training. On the third day after operation, the day of discharge respectively, the motor function and activity of daily living of the two groups were evaluated. The satisfaction of children′s parents on nursing was compared between the two groups.Results:The scores of motor function on the third day after operation and the day of discharge were (57.0 ± 6.8), (73.0 ± 5.4) points respectively in the observation group, and those in the control group were (55.0 ± 5.6), (65.6 ± 5.9) points. There were significant differences intra group comparison ( t=-9.26, -6.48, both P<0.05). The activity of daily living scores on the third day after operation and the day of discharge were (45.2 ± 5.9), (74.2 ± 8.3) points respectively in the observation group, and those in the control group were (44.0 ± 5.8), (60.2 ± 7.6) points. There were significant differences intra group comparison ( t=-14.33, -8.51, both P<0.05). There were significant differences between the scores of motor function and the ability of daily living scores on the day of discharge in the observation group and the control group ( t values were -4.65, -6.25, both P<0.05). The satisfaction of children′s parents on nursing in the observation group and the control group were 96.0% (24/25), 72.0% (18/25) respectively. There was significant difference between the two groups ( χ 2=8.78, P<0.05). Conclusions:Rehabilitation nursing based on guided education and training can improve motor function and ability of daily living in movement disorders school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection, promote the functional recovery of children after surgery and increase the satisfaction of children′s parents on nursing.