Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

Objective To evaluate the impact and clinical significance of NOD-like receptor pyrin domain-containing protein 3(NLRP3)in the activation of lipopolysaccharide(LPS)-induced BV2 microglia cells.Methods shRNA plasmids were devised for BV2 microglia transfection,and the most effective transfection segment was identified via RT-PCR and Western blot assays.NLRP3 expression in the cell line was detected by Western blotting,while light microscopy was used to observe morpho-logical alterations in BV2 cells transfected with NLRP3-shRNA.Furthermore,enzyme-linked immunosorbent assay(ELISA)was used to quantify levels of inflammatory cytokines IL-18,IL-1β,TNF-α and NO in cell supernatants.Immunofluorescence staining was used to observe the expression and localization of microglial activation markers iNOS,Arg-1,Iba1,and NLRP3.Results NLRP3 was highly expressed in LPS-induced BV2 cells.The Western blot result showed that the mRNA expression level was the lowest and transfection was the least effective in NLRP3-mus-727 group.Microscopic examination revealed a round or short spindle-shaped morphology in BV2 cells transfected with shNLRP3,which was akin to resting state cells in the blank control group.ELISA showed that pro-inflammatory mediators IL-18,IL-1β,TNF-α and NO levels were decreased in BV2 cells trans-fected with shNLRP3(all P＜0.05).Immunofluorescence staining indicated a relative decrease in iNOS and Iba1 expression,with an upregulation of Arg-1 in BV2 cells transfected with shNLRP3.Conclusion NLRP3 is highly expressed in LPS-induced BV2 cells.Inhibition of NLRP3 expression can suppress the inflammatory response of BV2 cells induced by LPS,promoting their polarization towards the M2 phenotype.

Objective:To explore the effect of different glucose concentrations on the uptake of 18F-FDG and the expression of glucose transport protein(Glut)-1 and Glut-3 in non-small cell lung cancer (NSCLC). Methods:NSCLC cell line A549 cells were cultured in DMEM medium with glucose concentrations of 3.9, 5.0, 6.1, 8.3 and 11.1 mmol/L respectively for 24 h. Then 3.7×10 4 Bq 18F-FDG was added into each group and γ counter was used to measure the radioactivity count 1 h later. Western blot was used to examine the expression of Glut-1 and Glut-3. One-way analysis of variance and Bonferroni test were used for data analysis. The correlation was analyzed by Pearson correlation analysis. Results:The 18F-FDG uptake rates in 3.9, 5.0, 6.1, 8.3 and 11.1 mmol/L groups were (4.89±0.83)%, (4.07±0.23)%, (3.66±0.29)%, (3.34±0.16)% and (3.29±0.24)%, respectively ( F=7.05, P=0.006). Compared with 3.9 mmol/L group, the 18F-FDG uptake rates in 8.3 and 11.1 mmol/L groups were reduced and differences were statistically significant ( P values: 0.013, 0.010), while there were no statistical differences between the other groups ( P values: 0.057-0.999). The relative expressions of Glut-1 and Glut-3 in each group were 1.17±0.10, 1.00±0.00, 0.84±0.07, 0.70±0.18, 0.61±0.16, and 1.14±0.05, 1.00±0.00, 0.86±0.12, 0.71±0.05, 0.40±0.06, respectively ( F values: 10.26 and 51.94, P values: 0.001, <0.001). Moreover, the 18F-FDG uptake rates were positively correlated with the expression of Glut-1 and Glut-3 ( r values: 0.775 and 0.744, both P=0.001). Conclusions:When the glucose concentration fluctuates within 3.9-11.1 mmol/L, the change of glucose will affect the 18F-FDG uptake rate and the expression of Glut-1 and Glut-3 in A549 cells. Moreover, the 18F-FDG uptake rate is related to the expressions of Glut-1 and Glut-3.

Objective:To investigate the distribution of 99Tc m-methylene diphosphonate (MDP) at different stages of bone injury repair. Methods:A total of 30 rabbit models of femur injury were established by the method of electric drill and perforation of femur. According to the different stages of bone injury repair (at 1, 2 and 3 week), rabbits were divided into group A, B and C ( n=10 each group). Femoral SPECT/CT imaging was performed on the last day of different stages of bone injury repair to obtain radioactivity counts in the region of interest (ROI) on the test side and control side and to calculate target/background ratio (T/B). The light intensity of 3 groups was analyzed by phosphor screen imaging and the distribution of 99Tc m-MDP in bone cells was observed by autoradiography. One-way analysis of variance and paired t test were used to analyze the data. Results:The T/B values of group A, B and C were 1.16±0.14, 1.39±0.23 and 1.18±0.10, respectively ( F=5.83, P<0.01). There were significant differences of the maximum radiation count between the test side (50.00±12.45, 59.50±12.83 and 55.10±9.26) and the control side (43.20±9.57, 50.00±12.30 and 44.30± 6.50) in group A, B and C ( t values: 3.24, 2.28 and 5.77, all P<0.05). There were significant differences in the light intensity of bone specimens in group A, B and C by phosphor screen imaging (37 324.67±6 481.50, 60 950.33±9 781.72 and 43 905.00±4 957.92; F=8.25, P=0.02). 99Tc m-MDP were deposited in both intracellular and extracellular during different stages of bone repair in osteocytes and osteoblasts under autoradiography. Conclusion:At different stages of bone injury repair, the concentration of 99Tc m-MDP is significantly distributed, suggesting that there are other ways of concentration mechanism of 99Tc m-MDP in bone tissue besides the chemical adsorption with hydroxyapatite.

Objective To investigate hand hygiene(HH) status among health care workers(HCWs) in municipal hospitals in Chongqing City, and provide the basis for making effective HH management strategies.Methods In April-June 2016, HH status among 111 HCWs in 24 municipal hospitals of this city were investigated through questionnaire survey, on-site observation, and hand surface sampling.Results All surveyed departments are installed special hand washing facilities, all surveyed HCWs were performed HH through hand-washing by running water.The proportion of HCWs' hand-washing by disinfectant was higher than six-step hand washing (73.87% [n=82] vs 37.84%[n=42], χ2=29.23, P<0.01);the implementation rate of HH before touching patient was higher than that after touching patients (99.10%[n=110] vs 89.19%[n=99], χ2=9.88, P<0.01).During the process of diagnosis and treatment activities, the maximal total number of bacteria on the surface of hand before and after HH were 475 CFU/cm2 and 85 CFU/cm2 respectively, hand surface colony count after HH was higher than before HH(P<0.01).Age, gender, department, and occupation are important factors influencing HH.The total number of bacteria on hand surface of nurses was higher than non-nurse HCWs, the total number of bacteria on hand surface of female, nurses, and HCWs in class I environment were all higher than male, non-nurse HCWs and HCWs in other types of environment, there were significant difference among the groups (all P<0.05).Qualified rates of HH of each group improved after hand washing, the total number of bacterial colony on hands of HCWs all decreased.Conclusion Hand washing facilities and HH efficacy are good in Chongqing municipal hospitals, however,HH compliance needs to be improved among HCWs aged≥35 years,male HCWs, HCWs in class III and IV environmental departments, as well as non-nurse HCWs.

Objective To study the protective effect of Xingnaojing combined with alprostadil on brain after acute ischemic stroke in rats.MethodsSixty patients with acute ischemic stroke were enrolled in zhejiang xin'an international hospital from March 2014 to March 2016.They were randomly divided into control group and treatment group, 30 cases in each group.The control group received conventional treatment plus alprostadil, the treatment group in the control group based on the combination of Xingnaojing treatment.Two groups of patients after treatment, are given nursing intervention, such as routine diet guidance, nutritional support, health education.The levels of serum oxidative stress (MDA), vascular endothelial growth factor (VEGF) and vascular endothelial growth factor (VEGF) were compared between the two groups before and after treatment.The levels of cerebral blood flow (CBFV) were recorded before and after treatment Observe the adverse reactions during treatment.ResultsAfter 14 days of treatment, the NIHSS score of the treatment group was lower than that of the control group and the ADL score was higher than that of the control group.The difference between the two groups was statistically significant (P<0.05).Before treatment, the oxidative stress indexes MDA and Hcy were no significant difference between the two groups.After treatment, the oxidative stress indexes MDA and Hcy were lower than the control group(P<0.05).Before treatment, the levels of VEGF and CBFV in the two groups were no significant difference between the two groups.After treatment, the levels of VEGF and CBFV in the two groups were significantly higher than those in the control group (P<0.05).The adverse reaction rate between the 2 groups was similar, and there was no significant adverse reaction, there was no significant difference between the two groups.ConclusionXingnaojing combined with alprostadil has a certain clinical effect on acute ischemic stroke, and has a good protective effect on brain tissue after reperfusion.

Background and purpose:The proportion of incidental thyroid cancer in PET imaging was sig-niifcantly increased with the wide application of18F-FDG PET/CT. The correlation between the glucose metabolism of thyroid incidental thyroid cancer and pathological changes is unclear. The objective of this study is to analyze the relationship between the glucose metabolism by18F-FDG PET/CT and tumor differentiation or lymph node metastasis in patients with incidental thyroid cancer.Methods:A total of 195 patients with focal FDG-avid thyroid incidentaloma during cancer evaluation were enrolled. Fifty-three patients were diagnosed as having thyroid cancer by the pathology. The SUVmax of thyroid cancer foci, lesion size, lesion number, and SUVmax of normal thyroid tissue were quantiifed. The tumor pathological grades and lymph node metastasis were analyzed by the pathology. The patients were broken down into 4 groups (G1, G2, G3 and G4) according to the tumor pathological grades and lymph node metastasis (differentiated thyroid cancer, non-differentiated thyroid cancer, without lymph node metastasis and with lymph node metastasis). The differences of glucose metabolism between G1and G2groups, G3 and G4 groups were analyzed.Results:Fifty-three foci were found by PET imaging. Sixty-two foci were found by the pathology (37 papillary thyroid carcinoma, 4 follicular thyroid carcinoma, 9 medullary thyroid carcinoma, 3 poorly differentiated thyroid cancer). The SUVmax in the normal thyroid tissue, G1and G2were 1.51±0.30, 4.25±1.70 and 6.34±2.45, respectively. The SUVmax in the G1and G2were signiifcantly higher than in the normal thyroid tissue (t=11.0,t=7.10,P<0.01). The SUVmax of G3was 4.77±2.15, and the SUVmax of G4group was 4.67±2.02. The differences between G1and G2groups, G3 and G4groups were not statisti-cally signiifcant (t=3.61,P>0.05;t=0.33,P=0.56).Conclusion:The differentiated incidental thyroid carcinoma and non-differentiated incidental thyroid carcinoma had high glucose metabolism, and there was no signiifcant difference in the levels of glucose metabolism in different differentiation degree and metastasis ability cancer.

Objective To compare the imaging features of 18F-FDG PET/CT in malignant pleural effusion (MPE) and tuberculosis pleural effusion (TPE).Methods From January 2012 to December 2014,93 patients (52 males,41 females,and average age (66.7±12.7) years) with unexplained pleural effusion who underwent 18F-FDG PET/CT were retrospectively studied.MPE and TPE were confirmed by histology,cytology or clinical follow-up.Lesion SUVmax and T/NT were calculated.Both lesion size and density (in Hounsfield units) of pleural abnormalities on CT images were measured.The difference of images between MPE and TPE was analyzed.Diagnostic efficiency of 18F-FDG PET/CT for detecting MPE and TPE were estimated.Two-sample t test and x2 test were used to analyze the data.Results The lesion SUVmax of 66 patients with MPE and 27 patients with TPE was higher than that in normal tissues (7.72±6.50,8.43±4.92;t=7.81 and 7.15,both P＜0.01),but there was no statistical difference between the TPE and MPE (t=0.56,P＞0.05).T/NT was not significantly different between MPE and TPE (5.40±4.29,5.20±2.73 respectively,t=-0.22,P＞0.05) either.The uptake features of 18F-FDG were different between MPE and TPE (x2=29.3,P＜0.01).When the nodular 18F-FDG uptake increase in pleura was taken as the malignancy,diffuse 18F-FDG uptake increase in pleura as tuberculosis,the sensitivity,specificity,positive predictive value,and negative predictive value of 18F-FDG PET/CT were 78.8％ (52/66),81.5％ (22/27),91.2％ (52/57),61.1％ (22/36),respectively,for differentiation of MPE from TPE.Primary cancers were found in 43 patients.When the nodular uptake increase in pleura,or primary cancer associated with increased pleural uptake found by PET was taken as the malignancy,the above mentioned parameters were95.5％(63/66),81.5％(22/27),92.6％(63/68),88.0％(22/25),respectively.Conclusions The 18FFDG uptake in MPE and TPE is higher than that in normal tissues.18F-FDG PET/CT is helpful in the search for the primary tumor of MPE.Qualitative method of 18F-FDG PET/CT has a good value in the differentiation of MPE from TPE.

Objective: To study the protective effect of baicalin on myocardial ischemia reperfusion injury and its correlation to myocardial cell autophagy in experimental rats. Methods: The animal models were established by intragastric infusion at 7 days prior operation in different groups.Atotal of 48 rats were divided into 4 groups: Sham operation group, Sham+baicalin group, Ischemia reperfusion (IR) group and Baicalin treatment group. n=12 in each group. Hemodynamics at different time points and myocardial infarction (MI) size at 45 min after reperfusion were recorded; protein expressions of LC3-II and autophagy-related Becl-1 at 30 min after reperfusion were examined by Westernblot analysis; the opening condition of mitochondrial membrane channel transition pore (mPTP) was detected by NAD+ content. Results: The MI sizes in Sham operation group and Sham+baicalin group were too small to compare; MI sizes in IR group and Baicalin treatment group were (41.32±1.85) % vs (23.30±1.60) %, P<0.001. Protein expressions of LC3-II in IR group and Baicalin treatment group were (1.051±0.005) and (0.863±0.009) which were both higher than Sham operation group (0.763±0.007), P<0.01;Becl-1 were (1.169±0.002) and (0.943±0.005) which were both higher than Sham operation group (0.647±0.014),P<0.01; LC3-II and in Becl-1 expressions in Baicalin treatment group were decreased than IR group, P<0.01. NAD+ contents (by nmol/mg) in IR group and Baicalin treatment group were (6.02±0.33) and (9.56±0.53) which were both lower than Sham operation group (11.28±0.37), P<0.001; NAD+ content in Baicalin treatment group was increased than IR group,P<0.01. Conclusion: Baicalin had no autophagy effect in normal myocardial cells, but it may decrease the MI size and reduce excessive autophagy in myocardial cells after IR which might be related to inhibiting mPTP opening.