Objective:To investigate the predictive value of 18F-fluorodeoxyglucose (FDG) PET/CT metabolic parameters for occult lymph node metastasis (OLM) in non-small cell lung cancer (NSCLC). Methods:A total of 183 patients (72 males, 111 females; age (61.5±8.4) years) who underwent 18F-FDG PET/CT and preoperatively diagnosed with clinical N0 stage (cN0) in Third Affiliated Hospital of Soochow University from January 2013 to December 2018 were retrospectively enrolled. All patients underwent anatomical pulmonary resection with systematic lymph node dissections within 3 weeks after 18F-FDG PET/CT examinations. According to the presence or absence of lymph node metastasis, patients were divided into OLM positive (OLM+ ) group and OLM negative (OLM-) group. Parameters of primary lesions, such as the maximum diameter (D max), tumor sites, morphological features, maximum standardized uptake value (SUV max), mean standardized uptake value (SUV mean), metabolic total volume (MTV), total lesion glycolysis (TLG), tumor SUV max to liver SUV mean (TLR max), tumor TLG to liver SUV mean (TLR TLG) were analyzed. Mann-Whitney U test and χ2 test were used to compare the parameters between groups. Multivariable logistic regression was used to analyze the independent risk factors for OLM. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic value of different parameters. Results:Among 183 patients, 25 (13.7%, 25/183) of them were diagnosed as OLM. In OLM+ group, 46 lymph nodes were pathologically positive for metastasis, including 15 N1 disease and 31 N2 disease. D max (2.9(2.3, 3.7) vs 2.3(1.7, 2.8) cm), lobulation ((76.0%(19/25) vs 37.3%(59/158)), SUV max (11.1(7.9, 17.7) vs 4.7(2.3, 9.2)), TLG (41.5(10.2, 91.1) vs 15.6(6.5, 23.8) ml), TLR max (4.7(3.5, 7.6) vs 2.1(0.9, 4.0)) and TLR TLG (18.1(5.0, 44.3) vs 6.1(3.0, 11.4) ml) of the primary lesions in OLM+ group were significantly higher than those in OLM-group ( z values: from -4.709 to -3.247, χ2=13.190, all P<0.05). Multivariable logistic regression analysis showed that TLR max (odds ratio ( OR)=15.145, 95% CI: 3.381-67.830, P<0.001) and D max ( OR=3.220, 95% CI: 1.192-8.701, P=0.021) were independent risk factors for OLM. TLR max yielded the highest area under curve (AUC; AUC=0.794) with the threshold of 3.12, and the sensitivity, specificity, accuracy, positive predictive value and negative predictive value for predicting OLM were 92.0%(23/25), 63.3%(100/158), 67.2%(123/183), 28.4%(23/81) and 98.0%(100/102), respectively. Conclusions:TLR max of tumor is the independent risk factor for OLM in NSCLC patients. TLR max can sensitively predict OLM preoperatively in patients with NSCLC.

Background Efficient and lowcost way to isolate keratocytes is helpful for research on cornea.Either relatively expensive or inefficient is the shortage of those means now applied,while raising the keratocytes through passage will change the phenotype of them quickly.Our aim is to approach the way getting keratocytes effectively utilizing modified two step enzymatic digestion by type I collagenase. Objective To evaluate the effect of isolating the bovine keratocytes utilizing two step enzymatic digestion and observe the morphological changes of the keratocytes during cultivation in vitro. Methods Keratocytes were isolated from bovine corneas using 0.5 mg/mL and 1 mg/mL type I collagenase digestion.The harvesting rate and viability rate of the primary keratocytes were evaluated.During the primary cultivation in vitro,the morphological changes of the keratocytes and their F-action distribution were observed.Results(1)The extracellular matrix of the bovine corneas were almost dissolved by the two step enzymatic digestion,followed the keratocytes completely isolated from the solid matrix.The amount of the harvested keratocytes from each cornea was(2.11±0.15)X106 on average while the viability rate was(91.69±3.73)% and the inoculation rate Was(81.20±1.25)%.(2)The primary keratocytes attached and spreaded out with dendritic and stellate morphology.After 3 days cultured,the branches of the keratocytes were contacting and formed networks.The F-actin detected by phalloidin binding showed a limited cortical localization. Conclusion (1)The method of two step enzymatic digestion can make the extracellular matrix of bovine cornea stroma completely degraded with the advantages in high efficiency of harvesting keratocytes and high cell viability and relatively simple manipulation. (2) The primary bovine keratocytes have dendritic morphology and with limited F-action distribute in cellular cortex.

Objective 　To evaluate the value of 99m　Tc-TRODAT-1 SPECT DAT imaging in the early diagnosing of Parkinsons disease (PD).Methods　 Eleven patients (9 PD and 2 possible PD) and eighteen healthy subjects matched by sex and age were studied with 99m　Tc-TRODAT-1 SPECT DAT imaging. Striatum specific uptake of 99mTc-TRODAT-1 was calculated according to the ratio of DAT uptake in striatum (ST) and cerebellum (CB). Results　In the hemi-Parkinsons disease group, the DAT specific uptake of 99mTc-TRODAT-1 was significantly lower (P＜0.01) in contralateral than in ipsilateral striatum to the clinically symptomatic side. There was significant decrease (p＜0.01) of striatum DAT uptake in patients with hemi-PD compared to the controls.Conclusions　99mTc-TRODAT-1 SPECT DAT imaging may help to confirm the diagnosis of PD at the early stage.

OBJECTIVETo explore the intervention of baicalin on signal transduction and activating transcription factor expression of ulcerative colitis (UC) patients.

METHODSRecruited were UC patients at Outpatient Department of Digestive Disease, Inpatient Department of Digestive Disease, Center for Digestive Endoscopy of College City Branch, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Southern Hospital affiliated to Southern Medical University from June 2010 to January 2011. They were assigned to the UC group (33 cases) and the diarrhea-predominant irritable bowel syndrome (IBS-D) group (30 cases). Another 30 healthy subjects were recruited as a healthy control group. Peripheral blood mononuclear cells (PBMCs) in vitro intervened by different concentrations baicalin were taken from UC patients. IL23R gene expressions in vitro intervened by different concentrations baicalin were detected using Q-PCR. Expressions of signal transducer and activator of transcription 4 (STAT4) , STAT6, phosphorylated-STAT4 (p-STAT4), and p-STAT6 were detected using Western blot. Serum levels of IFN-γ, IL-4, IL-6, and IL-10 were measured by ELISA. Effects of different concentrations baicalin on expressions of PBMCs, and levels of IFN-γ, IL-4, IL-10 of UC patients were also detected.

RESULTSCompared with the negative control group, 40 µmol baicalin obviously decreased IL23R gene expression of UC patients (P <0. 01). Compared with the healthy control group and the IBS-D group, p-STAT4/STAT4 ratios increased, p-STAT6/STAT6 ratios decreased, levels of IFN-γ, IL-4, IL-10 all increased in the US group (all P <0. 05). Compared with the negative control, 5 and 10 µmol baicalin groups, 20 and 40 moL baicalin obviously decreased p-STAT4/STAT4 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously increased p-STAT6/STAT6 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously lowered levels of IFN-γ and IL-4, and elevated IL-10 levels (all P <0. 05).

CONCLUSION40 µmoL baicalin could in vitro inhibit p-STAT4/STAT4 ratios, adjust p-STAT6/STAT6 ratios and related cytokines, thereby balancing the immunity and relieving inflammatory reactions of UC.

Activating Transcription Factors ; metabolism ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Blotting, Western ; Colitis, Ulcerative ; drug therapy ; metabolism ; Cytokines ; metabolism ; Flavonoids ; therapeutic use ; Humans ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; Leukocytes, Mononuclear ; Medicine, Chinese Traditional ; Phosphorylation ; STAT6 Transcription Factor ; metabolism ; Signal Transduction

OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation

Adolescent ; Child ; Collagen Type I ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Fibrillin-1 ; Fibrillins ; Heart Defects, Congenital ; metabolism ; pathology ; Humans ; Male ; Microfilament Proteins ; metabolism

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

OBJECTIVETo study the relationship between hRad17 mRNA expression and clinicopathologic factors and lymph node metastasis of gastric cancer, and to assess the significance of predicting the extent of lymph node metastasis and prognosis.

METHODShRad17 mRNA expression was examined in matched primary lesions, normal gastric mucosa and lymph node metastatic lesions among 52 gastric cancer patients by reverse transcription polymerase chain reaction (RT-PCR), polyacrylamide gel electrophoresis (PAGE) and silver stain with the relation between hRad17 mRNA expression and clinicopathologic factors analyzed. At the same time, hRad17 mRNA expressions in 5 gastric benign lesions and SGC7901 gastric carcinoma cell lines were also examined.

RESULTSThe primary tumor samples (88.4% positive) showed a significantly higher level of hRad17 expression compared with matched normal tissue (76.9% positive) (P = 0.014), so did the lymph node metastatic samples (94.2% positive) (P = 0.001). The hRad17 mRNA expression showed a low level in benign lesions, but very high in SGC7901 cell line. The hRad17 mRNA expression showed a higher level in patients with the number of lymph node metastasis above 15 than below 15 (P = 0.02), so did the diffused growth than the mass-like growth (P = 0.04).

CONCLUSIONThe method of PAGE and silver stain can improve the sensitivity of RT-PCR. The degree of lymph node metastasis and invasiveness of carcinoma cells are more serious in cases with hRad17 mRNA overexpression, and extensive lymph node dissection should be carried out for these patients. Examination of hRad17 expression by RT-PCR before surgery is indicated to arrive at an optimum treatment scheme and to estimate the prognosis.

Cell Cycle Proteins ; genetics ; Gastric Mucosa ; metabolism ; Humans ; Lymphatic Metastasis ; RNA, Messenger ; analysis ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo observe the efficacy of biased constitution patients treated by auricular point therapy on the basis of "preventive treatment" theory.

METHODSBy means of self-control, 1477 cases of biased constitution were regulated by auricular point sticking therapy or acupuncture, and follow-up and statistical analysis were conducted to observe the efficacy.

RESULTSThere were 322 markedly effective patients, 914 effective patients, 241 failed patients and the total effective rate was 83.7% (1236/1477).

CONCLUSIONThe comprehensive auricular treatment which is easy to operate and acceptable for patients has obvious effect in biased constitution patients. As a new direction of the "preventive treatment" theory, it can be promoted to be a new intervention way of health care.

Acupuncture Points ; Acupuncture Therapy ; Acupuncture, Ear ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Preventive Medicine ; Young Adult

Objective To investigate the myocardial damage in rats fed with corn from Keshan disease area added with bean puree. Methods Male Wistar rats were randomly divided into 3 groups according to their body weights, and fed with corn, corn from Keshan disease area added with bean puree, corn from non-endemic area. The GSH-Px activity of vena cardalis blood was examined in 1 and 3 months, rats were sacrificed after being fed for 6 months to examine the heart changes with HE stain. Results The three groups of GSH-Px activity were different in 1 and 3 months respectively(F=23.60,72.46, all P<0.01); GSH-Px activity was (181.58±22.15), (44.76±28.59)U/L in rats fed with corn, was (195.03±17.66), (30.38±3.35)U/L in those fed with corn added with bean puree from Keshan disease area, lower than the group fed with corn of non-endemic area[(340.90±95.42), (125.17±13.64)U/L, all P < 0.01]. But the difference of GSH-Px activity between simple corn group and corn adding bean puree groups of Keshan disease area was not obvious(P>0.05). Myocardial damage incidence of the three groups was 3/9,1/9,2/7. Difference among three groups did not have statistical significance(χ2=1.33, P> 0.05). Conclusions Only corn from Keshan disease area may induce myocardial damage pathology change. Adding bean puree into corn does not increase damage.