Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Objective To investigate the intervention effect and mechanism of LUO's Neiyi Prescription on the proliferation and angiogenesis of eutopic endometrium in rats with endometriosis(EMs)based on hypoxia-inducible factor 1A(HIF1A)/enhancer of zeste homolog 2(EZH2)/anthrax toxin receptor 2(ANTXR2)signaling pathway.Methods SD rats were randomly divided into sham operation group,model group,Danazol group(4.2 g·kg-1),low-dose LUO's Neiyi Prescription group and high-dose LUO's Neiyi Prescription group(15.74,31.48 g·kg-1),eight rats in each group.The rat model of EMs with qi stagnation and blood stasis syndrome was constructed by autologous endometrial transplantation combined with multi-factor intervention.Intragastric administration was given once a day for 28 consecutive days.The pathological changes of endometrial tissue were observed by HE staining.The expression levels of proliferating cell protein 67(Ki67)and platelet endothelial cell adhesion molecule 1(CD31)in endometrial tissue were detected by immunohistochemistry.The mRNA and protein expression levels of HIF1A,EZH2 and ANTXR2 in endometrial tissues were detected by qRT-PCR and Western Blot.The expression levels of YAP1,CD44 and β-catenin in endometrial tissues were detected by Western Blot.Results Compared with the sham operation group,the epithelial cells of the eutopic endometrium of the model group were thickened,the interstitial cells were arranged disorderly,and the inflammatory cells increased.The expression levels of Ki67 and CD31 in endometrial tissues were significantly increased(P＜0.01),the mRNA and protein expression levels of HIF1A and ANTXR2 were significantly increased(P＜0.01),while the mRNA and protein expression levels of EZH2 were significantly decreased(P＜0.01),and the protein expression levels of YAP1,CD44 and β-catenin were significantly increased(P＜0.01).Compared with the model group,the epithelial layer of the eutopic endometrial tissue of the rats in each administration group became thinner,the interstitial disorder and inflammatory infiltration were improved,and the levels of Ki67 and CD31 in the eutopic endometrial tissue were significantly decreased(P＜0.01).The mRNA and protein expression levels of HIF1A and ANTXR2 in the endometrium of rats in the Danazol group and the high-dose LUO's Neiyi Prescription group were significantly decreased(P＜0.05,P＜0.01),and the mRNA and protein expression levels of EZH2 were significantly increased(P＜0.05,P＜0.01).The protein expression levels of YAP1,CD44 and β-catenin in endometrial tissue of rats in each administration group were significantly decreased(P＜0.05,P＜0.01).Conclusion LUO's Neiyi Prescription can play a role in the treatment of EMs by inhibiting the proliferation and angiogenesis of eutopic endometrial cells,and its mechanism may be related to the inhibition of HIF1A/EZH2/ANTXR2 signaling pathway.

Platinum antitumor drugs are widely used for clinical treatment because of their unique antitumor mechanisms, significant antitumor effects, and broad antitumor spectrum. Yet shortcomings such as toxic side effects, drug resistance and cross-resistance of platinum-based drugs have limited their further application. Platinum-intercalator conjugates possess different antitumor mechanisms from those of classic platinum drugs, and have unique advantages in overcoming the disadvantages of classic platinum antitumor drugs. The platinum-intercalator conjugates can be classified into six categories based on the different DNA-intercalator: platinum-acridine, platinum-quinoline, platinum-indole, platinum-naphthalimides, platinum-anthraquinone and platinum-based antitumor agents containing other types of intercalating groups. This article summarizes the research progress of platinum-based antitumor drugs containing DNA insertion groups in recent years.

In order to solve the pathological grading of hepatocellular carcinomas (HCC) which depends on biopsy or surgical pathology invasively, a quantitative analysis method based on radiomics signature was proposed for pathological grading of HCC in non-contrast magnetic resonance imaging (MRI) images. The MRI images were integrated to predict clinical outcomes using 328 radiomics features, quantifying tumour image intensity, shape and text, which are extracted from lesion by manual segmentation. Least absolute shrinkage and selection operator (LASSO) were used to select the most-predictive radiomics features for the pathological grading. A radiomics signature, a clinical model, and a combined model were built. The association between the radiomics signature and HCC grading was explored. This quantitative analysis method was validated in 170 consecutive patients (training dataset: = 125; validation dataset, = 45), and cross-validation with receiver operating characteristic (ROC) analysis was performed and the area under the ROC curve (AUC) was employed as the prediction metric. Through the proposed method, AUC was 0.909 in training dataset and 0.800 in validation dataset, respectively. Overall, the prediction performances by radiomics features showed statistically significant correlations with pathological grading. The results showed that radiomics signature was developed to be a significant predictor for HCC pathological grading, which may serve as a noninvasive complementary tool for clinical doctors in determining the prognosis and therapeutic strategy for HCC.
Carcinoma, Hepatocellular ; diagnostic imaging ; Humans ; Liver Neoplasms ; diagnostic imaging ; Magnetic Resonance Imaging ; Neoplasm Grading ; methods ; ROC Curve

Objective To investigate the effect of acupuncture combined with antidepressant on behavioral changes and 5-hydroxytryptamine 1A and 2A receptor ( 5-HT1AR and 5-HT2AR) mRNA expression in nucleus raphes dorsalis (NRD) of sleep deprivation depression (SDD) rats. Methods Eight normal Sprague Dawley (SD) rats were caged together without any stimulus or treatment, serving as the normal group. Thirty-two SD rats were given solitary raise for 21 days, together with chronic unpredictable mild stress ( CUMS) and rapid eye movement ( REM) sleep deprivation to establish the model. Twenty-eight rats completed the modeling successfully, and were divided into model group, acupuncture group, medicine group, and combination group, 7 rats in each group. Rats in the model group were given modeling treatment but without any intervention. Rats in the acupuncture group were given acupuncture on acupoint Yintang, Shenting and bilateral Taichong, acupuncture stimulation lasting 15 s every 10 min and acupuncture retention for 20 min in each day. Rats in the medicine group were given gastric gavage of Zoloft solution (0.83 mg·kg-1·d-1) and Alprazolam solution (0.067 mg· kg-1·d-1) , and rats in the combination group were given both acupuncture and medicine intervention. The intervention for the three intervention groups lasted for 7 days. Results Compared with the normal group, body weight, Open-field scores and sucrose preference of the model rats were significantly decreased on modeling day 7, 14, 21 (P<0.05). On modeling day 28, body weight and Open-field scores of each intervention group were significantly increased compared with those of the model group (P<0.05), and Open-field scores of combination group differed from those of acupuncture group and medicine group ( P<0.05). Only sucrose preference of combination group was improved significantly compared with the model group (P<0.05). Compared with the model group, NRD 5-HT1AR mRNA expression level was increased in the combination group (P<0.05), and NRD 5-HT2AR mRNA expression level was down-regulated in all of the three intervention groups ( P<0.05). Conclusion Acupuncture and antidepressant treatment covering 7 days can evoke rapid effect on SDD rats, which can improve the slow effect of antidepressant.

OBJECTIVETo establish an in vitro cell model for investigating hepatic steatosis in non-alcoholic fatty liver disease.

METHODSL-02 cells cultured in 1640 containing 10% fetal bovine serum were divided into control group and model group. At 70%-80% confluency, L-02 cells in the model group were exposed to a long-chain mixture of free fatty acids (FFA, oleate and palmitate ) for 24 h, and cells in control group were treated with fresh medium. Lipid droplets in the cells were observed and total lipid content was determined with Oil Red O staining. The morphology of lipid droplets, trilyceride level, malonaldehyde content and cell apoptosis rate were evaluated to verify the cell model, and the effect of Huganqingzhi tablet on the lipid droplets was observed.

RESULTSA large number of lipid droplets were found in the cell model, which showed markedly increased level of triglyceride without significant changes of malonadehyde content or cell apoptosis rate. Intervention with two doses of Huganqingzhi tablet significantly decreased the number of lipid droplets and trilyceride content in the cell model.

CONCLUSIONhepatic steatosis L-02 cell model can be established by long-chain mixture of free fatty acids (oleate:spalmitate=2:1) for therapeutic drug studies.

Apoptosis ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Fatty Acids, Nonesterified ; chemistry ; Fatty Liver ; Humans ; Malondialdehyde ; analysis ; Non-alcoholic Fatty Liver Disease ; Triglycerides ; analysis

Objective To establish an in vitro cell model for investigating hepatic steatosis in non-alcoholic fatty liver disease. Methods L-02 cells cultured in 1640 containing 10%fetal bovine serum were divided into control group and model group. At 70%-80%confluency, L-02 cells in the model group were exposed to a long-chain mixture of free fatty acids (FFA, oleate and palmitate ) for 24 h, and cells in control group were treated with fresh medium. Lipid droplets in the cells were observed and total lipid content was determined with Oil Red O staining. The morphology of lipid droplets, trilyceride level, malonaldehyde content and cell apoptosis rate were evaluated to verify the cell model, and the effect of Huganqingzhi tablet on the lipid droplets was observed. Results A large number of lipid droplets were found in the cell model, which showed markedly increased level of triglyceride without significant changes of malonadehyde content or cell apoptosis rate. Intervention with two doses of Huganqingzhi tablet significantly decreased the number of lipid droplets and trilyceride content in the cell model. Conclusion hepatic steatosis L-02 cell model can be established by long-chain mixture of free fatty acids (oleate∶spalmitate=2∶1) for therapeutic drug studies.

Objective To establish an in vitro cell model for investigating hepatic steatosis in non-alcoholic fatty liver disease. Methods L-02 cells cultured in 1640 containing 10%fetal bovine serum were divided into control group and model group. At 70%-80%confluency, L-02 cells in the model group were exposed to a long-chain mixture of free fatty acids (FFA, oleate and palmitate ) for 24 h, and cells in control group were treated with fresh medium. Lipid droplets in the cells were observed and total lipid content was determined with Oil Red O staining. The morphology of lipid droplets, trilyceride level, malonaldehyde content and cell apoptosis rate were evaluated to verify the cell model, and the effect of Huganqingzhi tablet on the lipid droplets was observed. Results A large number of lipid droplets were found in the cell model, which showed markedly increased level of triglyceride without significant changes of malonadehyde content or cell apoptosis rate. Intervention with two doses of Huganqingzhi tablet significantly decreased the number of lipid droplets and trilyceride content in the cell model. Conclusion hepatic steatosis L-02 cell model can be established by long-chain mixture of free fatty acids (oleate∶spalmitate=2∶1) for therapeutic drug studies.

Objective To observe the difference between skeletal age of hand and wrist and chronological age and explore the reliability of CHN radiographic atlas method to assess the skeletal age of hand and wrist in children and adolescent.Methods Total 1397 healthy children (666 boys,731 girls;age range,1.0-18.0 years old) with hand and wrist injury from 2007 to 2011 were selected.Forty groups (n =20 for boys and girls,respectively) were classified according to CHN radiographic atlas method.The radiographs of hand and wrist were assessed by CHN radiographic atlas method,the relations between skeletal age and chronological age were investigated by using Wilcoxon signed ranks test.Results According to the CHN radiographic atlas method,the difference in 1.0 to 3.9 years old,7.0 to 7.9 years old and 9.0 to 15.9 years old boy groups between skeletal age and the chronological age had statistical significance (P ＜0.05) ; the difference in 1.0 to 2.9 years old,8.0 to 11.9 years old,12.6 to 14.9 years old,and 17.0 to 18.0 years old girl groups between skeletal age and the chronological age had statistical significance (P ＜ 0.05).Besides,these skeletal age was higher than the chronological age.Conclusions Skeletal age assessed by the CHN radiographic atlas method in a majority of age groups was higher than chronological age.It should be cautious to estimate the contemporary Chinese children skeletal age of hand and wrist when using the CHN radiographic atlas method.