Objective:To investigate the expression of DARS2 and its clinical significance in colorectal cancer.Methods:In this study, bioinformatics tools, especially gene expression profile interactive analysis 2 (GEPIA2), were used to conduct an in-depth analysis of DARS2 expression in colorectal cancer tissues. Immunohistochemical staining was carried out in 108 colorectal cancer specimens and 30 normal colorectal tissues obtained from the First Affiliated Hospital of Nanchang University, Nanchang, China. Colorectal cancer cell lines (HCT116 and SW480) were transfected with small interfering RNA (siRNA) and DARS2 overexpression plasmid to examine the effects of DARS2 knockdown and overexpression on cell function. To assess the effects on cell function, CCK8 and transwell migration assays were used to assess proliferation and cell motility, respectively. Additionally, protein immunoblotting was employed to scrutinize the expression of proteins associated with the epithelial-mesenchymal transition of colorectal cancer cells.Results:DARS2 exhibited a pronounced upregulation in expression within colorectal cancer tissues compared to their normal epithelial counterparts. Furthermore, DARS2 expression was higher in colorectal cancer of stage Ⅲ-Ⅳ than those of stage Ⅰ-Ⅱ, exhibiting a significant correlation with N staging, M staging, and pathological staging ( P<0.05). Kaplan-Meier analyses showed a decreased overall survival rate in colorectal cancer with DARS2 expression compared to those without DARS2 expression ( P<0.05). In the siRNA transfection group, there was a significant reduction in cell proliferation and migration ( P<0.01 and P<0.05, respectively). Conversely, the transfection of DARS2 overexpression plasmids substantially increased both cell proliferation and migration ( P<0.05). Additionally, immunoblotting revealed that DARS2 knockdown led to an upregulation of E-cadherin expression and a downregulation of N-cadherin and vimentin expression. In contrast, DARS2 overexpression resulted in increased N-cadherin and vimentin expression, coupled with reduction in E-cadherin expression. Conclusions:There is a strong association between DARS2 expression and colorectal cancer progression. Silencing DARS2 inhibits cell proliferation and migration, exerting a discernible influence on the epithelial-mesenchymal transition process.

Objective: To investigate the significance of Twist2 in glioma and whether it is involved in the malignant transformation of glioma by epithelial-mesenchymal transition (EMT). Methods: Using immunohistochemical method detected the expression level of Twist2 in 60 cases of gliomas (including WHO grades Ⅱ, Ⅲ and Ⅳ, each for 20 cases) and 20 cases of non-tumor brain tissues. Real-time fluorescence quantitative PCR and Western blot were used to detect the expression level of Twist2 mRNA and protein in 61 cases of fresh glioma tissue (WHO grade Ⅱ 16 cases, Ⅲ 21 cases, Ⅳ 24 cases) and 12 cases of adjacent tissues, and the expression levels of E-cadherin, N-cadherin and vimentin were also investigated in fresh glioma tissue. Results: Immunohistochemistry results showed that the percentages of Twist2 expression in glioma was 90%(54/60) compared with 30%(6/20) in non-tumor brain tissues(P<0.01). The percentages of Twist2 expression were 75% (15/20), 95% (19/20), and 100% (20/20) in the WHO gradesⅡ, Ⅲ and Ⅳ gliomas, respectively. WHO grades Ⅳ and Ⅲ were significantly higher than that of WHO grade Ⅱ (P<0.01). There was no significant difference between WHO grade Ⅳand WHO Ⅲ glioma (P>0.05). Real-time fluorescence quantitative PCR and Western blot showed that the expression level of Twist 2 in gliomas was significantly higher than that in para-cancerous tissues (P<0.01), and those in WHO grades Ⅳ and Ⅲ gliomas were significantly higher than that in WHO grade Ⅱ glioma (P<0.01). There was no significant difference between WHO grade Ⅳand grade Ⅲ glioma (P>0.05). Detection of key protein expression in EMT by Western blot displayed that the expression of E-cadherin was negatively associated with Twist2 in glioma (r=-0.972, P<0.01). The expression of N-cadherin and vimentin was positively associated with Twist2 in glioma(r=0.971, P<0.01; r=0.968, P<0.01). Conclusions The expression of Twist2 in human glioma is positively correlated with the malignant grade of glioma, which may be involved in the malignant progression of glioma by EMT.

Objective To discuss pathological features of three cases of uterine primary NHL(non Hodgkin′s lymphoma).Methods The clinical and pathological data of 3 cases of NHL were collected.Immunohistochemical method was used to label the tumor cells.Their histological types were classified by the latest WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues,and then differential diagnosis with other uterus diseases was offered.Results Three cases were presented with abnormal vaginal bleeding or ejecting as the first symptom,which was easily misdiagnosed as common diseases of the uterus in preliminary clinical and pathological diagnosis.Histopathology of general uterus specimens demonstrated that case one expressed CD3,CD56,TiA-1 and so on,and detected EBV virus cells expression,which supported the diagnosis of NK/T cell lymphoma.Another two cases expressed B cell-specific markers(CD20) and so on,and these tumor cells volume was 3 times larger than that of lymphocyte.These tumor cells had high proliferation index and supported diffuse large B-cell lymphoma diagnosis.Conclusion The tumor cells are like lymphocytes.We infiltrate endometria and myometrium,which is difficult to identify from endometria stromal sarcoma and other tumors,and need to use immunohistochemistry(IHC) for definitive diagnosis and differential diagnosis.

Objective To evaluate the value of fetal intelligent navigation echocardiography with color flow imaging ( 5D Heart Color) in the display of key diagnostic elements in basic fetal echocardiographic views . Methods The heart volumes datasets of 220 fetuses were collected and the offline analysis was conducted . Pearson correlations was used to compare the detection rate of blood flow element between 5D Heart Color software and color Doppler flow imaging(CDFI) in basic fetal echocardiographic views ,and the detection rate of blood flow between normal fetuses ( control group ) and fetuses with cardiovascular abnormalities( case group) . Bland-Altman analysis was used to prove consistency and reliability of the results . Results Totally 384 fetal heart volume datasets were collected from 220 fetuses ,and 317 volume datasets of 200 cases could be used for the analysis ,with the acquiring success rate of 82 .6％ . 5D Heart Color software and CDFI showed a close correlation in displaying key cardiovascular blood flow information in key fetal echocardiographic diagnosis sections ( r =0 .88 , P <0 .05) ;The detection rates of blood flow elements in control group and case group were 91 .9％ and 87 .3％ ,respectively , with an average of 89 .6％( r = 0 .9 , P < 0 .05 ) . Bland-Altman analysis showed close consistency between inter- and intra-observations in the detection rates of blood flow elements ,and their 95％ confidence intervals were ( -0 .1/+0 .09) and ( -0 .09/+0 .1) ,respectively ,and the points outside the 95％ confidence interval were 5 .4％(17/231) and 5 .7％ (18/231) ,respectively . Conclusions 5D Heart Color software shows similar usefulness in the displaying key blood flow elements ,comparing with CDFI , demonstrating higher reliability and repeatability in displaying fetal echocardiographic diagnosis views and diagnostic blood flow elements .

The tumor multidrug resistance reversal effect of NPB304, a novel taxane, was studied. MTT assay was used to determine the IC50 of chemotherapy drugs. Western blotting assay was applied to analyze the expression of P-glycoprotein (P-gp). The effect of compounds on the P-gp function and P-gp ATPase activity was determined by rhodamine 123 (Rh123) accumulation assay and analysis kit, respectively. Molecular docking was employed to predict the binding force between compounds and P-gp. Transmembrane transport of NPB304 was analyzed using MDCK II and MDR1-MDCK II cell model. NPB304 displayed multidrug resistance reversal effect on KBV cells and MCF-7/paclitaxel cells, NPB304 collaborative with P-glycoprotein (P-gp) inhibitors verapamil enhanced the reversal activity, specifically, 10 μmol x L(-1) verapamil in combination with paclitaxel reversed resistance by 56.5-fold, while combined with NPB304 increased the reversal fold; NPB304 synergistically increased Rh123 accumulation in the resistant cells when combined with verapamil, and NPB304 at 0-1 μmol x L(-1) enhanced the ATPase activity activated by verapamil was observed. NPB304 existed the hydrophobic interactions with the TM regions of P-gp, and the binding force between NPB304 and the A chain of the TM region was stronger. P-gp ATPase activity assay demonstrated NPB304 at lower concentrations (0-1.5 μmol x L(-1)) could activate the P-gp ATPase, playing a role on inhibition of P-gp function. However, NPB304 did not have an obvious feature of P-gp substrate. NPB304 exerted itself and synergy with verapamil activity on reversing tumor resistance via inhibiting the P-gp function.

Objective To study the expression and its clinical significance of Oct4 in prostate cancer.Methods The Oct4 mRNA expression levels in 2 normal prostate (NP) samples and 8 prostate cancer (Pca) samples were determined by reverse transcription polymerase chain reaction (RT-PCR) analysis.The Oct4 protein expression levels in 10 NP samples,10 benign prostate hyperplasia (BPH) samples and 45 Pca samples were detected by immunohisochemical analysis.Results The relative expression levels of Oct4 mRNA were 0.91,0.76,0.66,0.63,0.55,0.50,0.48,0.42 in 8 Pca samples and 0 in 2 NP samples,respectively.The positive rates of Oct4 protein expression was higher in Pca tissues than in NP and BPH tissues (91.1％ vs.0.0％,91.1％ vs.0.0％,both P＜0.05).The level of Oct4 protein was increased with higher hostologic grade and clinical stage of prostate cancer.The strong positive rates of Oct4 expression were 72.7％ and 30.0％ in Pca tissues with and without lymph node metastasis respectively,and there was a significant difference in the strong positive rate between them (P＜0.05).Conclusions The Oct4 expression is closely related to the differentiation and invasion of tumor.Oct4 might be an important index for the prognosis of prostate cancer.

BACKGROUND:Both acupuncture and moxibustion and drug ale effective methods to treat simple obesity.Whether electroacupuncture is different from sibutramine to affect adipocyte products in obese organisms?OBJECTIVE:To observe the different effects of electroacupuncture and sibutramine on adipocyte produces in obesity rats and to investigate the mechanism of weight reduction by acupuncture and moxibustion.DESIGN:Layering-randomized controlled study.SETTING:Laboratory of Acupuncture and Pharmacology.Nanjing University of Chinese Medicine.MATERIALS:12045-day-old male SD rats were provided by Shanghai Experimental Animal Center.The experiment was in accordance with animal ethics standards.Sibutramine hyrdrochloride capsule was made in Fuling Pharmaceutical Factory of Taiji Group(batch number:X20010279).Electro-acupuncture apparatus(LH402A acupoint neutoelectrical stimulation equipment)was made in Beijing Huawei Chanye Developing Company.METHODS:This study was performed at the Laboratory of Acupuncture and Pharmacology,and Expefimental Animal Center,Nanjing University of Chinese Medicine from October 2004 to November 2006.①100 male SD rats underwent high-fat diet.Fifty-four alimentary obesity rats were randomly divided into electroacupuncture group,sibutramine group and obesity group.wim 18 rats in each group.Another 18 rats were regarded as a normal group.②Rats in the electroacupuncture group were given electrical stimulation(2/15 Hz,4 mA)at lateral Housanli and Neiting acupoints,contralateral alternation every other day.Rats were fixed with self-made binding instrument,and rats maintained a consciousness but not an intense struggle.The electricai stimulation lasted for 15 minutes every day,one day was an interval every 6 days.and the electroacupuncture was given for 49 days and 42 times in total.Rats in the sibutramine group were ingragastrically administrated with 0.2 g/L sibutramine hydrochloride(2.0 mg/kg/d).Rats in the control group and obesity groups were fixed the same as electroacupuncture group but not given electrical stimulation.MAIN OUTCoME MEASURES:①Obese marker:To detect body mass,body length,weight reduction rate and body mass/body fat mass,and to measure total cholesterol(TC),high density lipoprotein cholesterol(HDL-C)and triacylglycerol (TG)with biochemical chromatometry;②insulin sensitivity index:To measure fasting blood glucose,insulin and insulin sensitivity index with enzyme-linked immunosorbent assay;③adipocyte secreting regulatory substance level:To measure serum resistin,leptin,adiponectin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and its soluble receptor level with enzyme-linked immunosorbent assay.RESULTS:Among 120 rats,54 alimentary obesity rat models and 18 normal rats were included in the final analysis.①Obesemarker:Bodymass,bodyfatmass,cholesterol and TG levels in the obesity group were significantly higher than in the normal group(P＜0.01).Body mass,body fat mass,body mass/body fat mass,cholesterol and TG levels in the electroacupuncture group were significantly lower than in the obesity group(P＜0.01).TG level in the sibutramine group was significantly lower than in the obesity group(P＜0.01).Weight reduction rate in the electroacupuncture group was significantly higher than in the sibutramine group(P＜0.01).Body mass/body fat mass in the electroacupuncture group was significantly lower than in the sibutramine group(P＜0.01).②Insulin sensitivity:Blood glucose and serum insulin levels in the obesity group were significantly higher than in the normal group,and insulin sensitivity index was significantly lower than in the normal group(P＜0.01).Blood glucose and insulin levels in the electroacupuncture and sibutramine groups were significantly lower than in the obesity group,but insulin sensitivity index was significantly higher than in the obesity group(P＜0.01).③Adipocyte secreting regulatory substance level:Adiponectin in the electroacupuncture group was significantly higher than in the obesity group (P＜0.01).Resistin and TNF-αin the electroacupuncture group was significantly lower than in the obesity grollp(P＜0.01).Resistin level in the sibutramine group was significantly lower than in the obesity group(P＜0.01).CONCLUSION:The effect of electroacupuncture on abnormally secretary products of adipose tissues in obesity rats is superior to the effect of sibutramine.This may be one of important mechanisms of weight reduction by acupuncture and moxibustion,and prevention and cure of obesity and its related diseases.

Objective:To investigate if acupuncture has an inhibitory effect on fatty toxicity and its possible mechanism. Methods: Thirty-four patients with simple obesity were clinically treated with acupuncture for 3 courses. Before and after treatment, body mass index(BMI) and fat percentage(F%) were determined, and serum insulin and adiponectin were measured by enzyme linked immunoassay, blood lipid and sugar by biochemical colorimetry and eight kinds of free fatty acids(FFAs) by high performance liquid chromatography. Twenty normal persons were selected as a control. Results:The total efficacy rate was 88.2%. Acupuncture could increase the decreased insulin sensitivity index(ISI) in the patients(t=-5.103, P=0.000). The correlation of a decrease in F% with an increase in unsaturated fatty acid after acupuncture was of significance(r=0.402, P=0.019) and its correlation with an increase in the ratio between unsaturated fatty acid and saturated fatty acid was also of significance(r=0.348, P=0.044). The correlation between increases in high density lipoprotein and in eicosapentaenoic acid was of significance(r=0.352, P＝0.041). The correlation between a decrease in BMI and an increase in high density lipoprotein was of significance(r=0.357, P=0.038). Adiponectin level significantly rose after acupuncture (compared with pre-treatment, P=0.000). The correlation between a decrease in FFAs and an increase in adiponectin was of significance(r=-0.349, P=0.043). Conclusion: Acupuncture can lower the levels of free fatty acids in the patients and increase their sensitivity to insulin to inhibit the fatty toxicity. The inhibitory effect of acupuncture on fatty toxicity is somewhat related to a rise in adiponectin level.

BACKGROUND: The therapeutic effects of acupuncture on weight loss have been verified. How does the central neural system act on weight loss with acupuncture?What relationships are there between the central regulation and peripheral action?OBJECTIVE: To study the effect of acupuncture on feeding center of hypothalamus in obese cases.DESIGN: Randomized and controlled study in which the experimental animals were taken as the subjects.SETTING: Clinical medical college of a university of traditional chinese medicine.MATERIALS: The experiment was performed in Key Laboratory of Acupuncture and Moxibustion in Nanjing University of Traditional Chinese Medicine from June 1998 to January 2000. Fifty SD male rats were employed and randomized into acupuncture group, obese group and normal group.METHODS: The experimental obese model was prepared by feeding SD rats with high-fat forage. Microelectrode record of nerve cell and cerebral stereotaxic technique were applied, by the treatment with acupuncture on experimental obese rats, the electric activity (Hz) of nerve cell was observed at unit time in hunger center of lateral hypothalamus area(LHA) and satiety center of ventral medium nucleus of hypothalamus(VMH).MAIN OUTCOME MEASURES: Mass changes at the same stage before and after acupuncture in rats of every group and discharge frequency in LHA and VMH in rats of every group.RESULTS: Acupuncture reduced remarkably the excitability in LHA, the values of which in acupuncture, obese and normal groups were(9.4 ± 3.7),(21.4 ± 6. 8), (9. 1 ± 5.2) Hz respectively( P < 0. 01 ); improved electric activity frequency in VMH, the values of which in 3 groups were (21.1 ± 4. 3), (5.0 ± 1.3), ( 14. 5 ± 2. 2)Hz, respectively( P < 0.01 ), inhibited excessive appetite of obese rats and reduced calorie intake so as to achieve weight loss.CONCLUSION: The regulation of acupuncture on central neural nuclei in obese animals is one of the important mechanisms on weight loss.

Aim To explore the possible anti-inflammation mechanism of paeonol by investigating its effects on the MMP-9 mRNA expression and cytokines production in human dermal fibroblasts induced by TNF-?.Methods The effect of paneonol on the expression of MMP-9 mRNA was detected by semi-quantitative RT-PCR.The modulation of paneonol on the production of IL-1?,IL-6 and IL-8 in fibroblasts was measured by ELISA.Results MMP-9 hardly expressed in human dermal fibroblast.The results also showed that TNF-? significantly induced the expression of MMP-9 in fibroblasts and at the same time paeonol inhibited the expression of MMP-9.TNF-? stimulated the production of IL-1? and IL-8 in fibroblasts,while 10~100 mg?L-1 paeonol inhibited TNF-?-induced IL-1? and IL-8 production in fibroblasts but had nothing to do with the production of IL-6.Conclusions Paeonol can inhibit the expression of MMP-9 mRNA,IL-1? and IL-8 induced by TNF-?.