Accurate Measurement of Agatston Score Using kVp-Independent Reconstruction Algorithm for Ultra-High-Pitch Sn150 kVp CT

Objective@#To investigate the accuracy of the Agatston score obtained with the ultra-high-pitch (UHP) acquisition mode using tin-filter spectral shaping (Sn150 kVp) and a kVp-independent reconstruction algorithm to reduce the radiation dose. @*Materials and Methods@#This prospective study included 114 patients (mean ± standard deviation, 60.3 ± 9.8 years; 74 male) who underwent a standard 120 kVp scan and an additional UHP Sn150 kVp scan for coronary artery calcification scoring (CACS). These two datasets were reconstructed using a standard reconstruction algorithm (120 kVp + Qr36d, protocol A; Sn150 kVp + Qr36d, protocol B). In addition, the Sn150 kVp dataset was reconstructed using a kVp-independent reconstruction algorithm (Sn150 kVp + Sa36d, protocol C). The Agatston scores for protocols A and B, as well as protocols A and C, were compared.The agreement between the scores was assessed using the intraclass correlation coefficient (ICC) and the Bland–Altman plot. The radiation doses for the 120 kVp and UHP Sn150 kVp acquisition modes were also compared. @*Results@#No significant difference was observed in the Agatston score for protocols A (median, 63.05; interquartile range [IQR], 0–232.28) and C (median, 60.25; IQR, 0–195.20) (p = 0.060). The mean difference in the Agatston score for protocols A and C was relatively small (-7.82) and with the limits of agreement from -65.20 to 49.56 (ICC = 0.997). The Agatston score for protocol B (median, 34.85; IQR, 0–120.73) was significantly underestimated compared with that for protocol A (p < 0.001). The UHP Sn150 kVp mode facilitated an effective radiation dose reduction by approximately 30% (0.58 vs. 0.82 mSv, p < 0.001) from that associated with the standard 120 kVp mode. @*Conclusion@#The Agatston scores for CACS with the UHP Sn150 kVp mode with a kVp-independent reconstruction algorithm and the standard 120 kVp demonstrated excellent agreement with a small mean difference and narrow agreement limits. The UHP Sn150 kVp mode allowed a significant reduction in the radiation dose.

Comparative Analysis of the Components of Volatile Oil in Citrus medica from Different Producing Areas / 中国药房

OBJECTIVE：To compare th e difference of the components of volatile oil in Citrus medica from different producing areas. METHODS ：The volatile oil of C. medica from 10 different producing areas was extracted with steam distillation ，and the yield was calculated. The components of the volatile oil of C. medica from different producing areas were analyzed by GC-MS. The compounds were retrieved from NIST 14.L mass spectrum database and identified. Relative mass fraction of chemical component was determined by peak area normalization method. Cluster analysis of samples were performed by using SPSS 20.0 software. RESULTS：The yields of volatile oil of C. medica from 10 different producing areas were 0.10％-1.75％，among which sample from Qianwei county in Leshan city of Sichuan province was the highest （1.75％）. A total of 66 components were identified in the volatile oil of C. medica from different producing areas ，with a relative molecular weight of 126.20-392.66. The majority was C 10 and C 15 compounds；isomers with relative molecular weight of 136，154 took up the great proportion ，which were mainly cycloalkane monoterpenes. There were 12 common components in the volatile oil of C. medica from different areas ，which were limonene（24.90％），terpinene（14.71％），（－）-4 terpineol（2.88％），citral（2.33％），α-myrrhene（2.33％），geraniol（1.52％）， α-pinene（1.37％），trans bergamot olene （1.16％），isoterpinene（1.13％），methyl palmitate （1.12％），linalool（1.09％）and geranyl acetate（1.04％）according to relative mass fraction ；8 of them were monoterpenes ，2 were sesquiterpenes and 2 were esters. There were 4 categories of C. medica from different producing areas ，i.e. S 2，S4，S6 clustered into one ；S1，S3，S7，S8 clustered into one ； S5 and S 10 clustered into one ；S9 as one . CONCLUSIONS ： There are some difference of the components in volatile oil of medica from different producing areas ，and the content of the same component also has great difference in the volatile oil of C. medica from different producing areas.

Effects of endurance exercise on synaptic plasticity in cerebral cortex of aged rats and related regulatory mechanism / 中国应用生理学杂志

OBJECTIVE@#To understand and analyze the rules of endurance exercise on the cerebral cortex adaptive mechanism in aged rats.@*METHODS@#In this study, 3-month-old (n=20), 13-month-old (n=24) and 23-month-old (n=24) specific-pathogen free (SPF) male Sprague-Dawley Rat (SD) rats were divided into young (Y-SED), middle-aged (M-SED) and old-aged (O-SED) sedentary control group, and the corresponding Y-EX, M-EX and O-EX in the endurance exercise runner group. The 10-weeks of regular moderate-intensity aerobic exercise intervention were carried out in the endurance exercise runner group. The exercise mode is treadmill exercise (slope 0), and the exercise intensity gradually increases from 60%～65% of the maximum oxygen consumption (V·O) to 70%～75%, and the exercise time is 10 weeks. Hematoxylin and eosin (HE) staining was used to detect age-related morphological changes. The expressions of superoxide dismutase(SOD) and brain-derived neurotrophic factor (BDNF) and the expressions of synapsin 1 (SYN1) and Ca/calmodulin- dependent protein kinases IIα (CaMK IIα) / AMP-activated protein kinase α1(AMPKα1) / mammalian target of rapamycin (mTOR) pathway -related genes were detected.@*RESULTS@#The cerebral cortex structure of the rats in each group showed age-related aging changes, the expression of SOD in the cortex showed a gradual decline, the expression of BDNF showed an age-increasing trend, and the expression levels of SYN1 and CaMK IIα were increased with age. The changes in AMPKα1 and SirT2 and IP3R, AKT1 and mTOR mRNA levels were increased slightly in middle-aged rats and decreased in aged rats. Compared with the rats in each sedentary control group, the nucleus of the cerebral cortex was tightly arranged and the number of nuclei observed under the microscope was increased significantly in each exercise group. Exercise promoted the expressions of SOD, BDNF and synaptophysin SYN1 in the cortex of rats, and the expression levels of SOD and BDNF in aged rats were up-regulated significantly (P＜ 0.01). The expression level of SYN1 in rats was up-regulated significantly (P＜0.05) in the young and aged rats. The expression of CaMK IIα in the cortex of middle-aged and aged rats was up-regulated (P＜0.01), while the expression level of CaMK IIα in young rats was down-regulated (P＜0.01). Exercise could up-regulate the expression level of AMPKα1 in the cortex of young rats (P＜ 0.05), but not in middle-aged and old-age rats. Exercise could up-regulate the expression of SirT2 in the cortex of rats in all age groups (P＜0.05). Exercise up-regulated the expression of phosphoinositide 3-kinase (IP3R)/ protein kinase B 1(AKT1) /mTOR in the cortex of rats, among which young IP3R was significantly up-regulated (P＜0.01) in the young group, mTOR was significantly up-regulated in young and middle-aged group (P＜0.01), and mTOR was also significantly up-regulated in the aged group (P＜0.05).@*CONCLUSION@#Endurance exercise up-regulates BDNF expression, regulates CaMKIIα signaling, activates AMPK signaling pathway and IP3R / AKT1 / mTOR signaling pathway, and improves synaptic plasticity in the cortex.

Study on the Separation and Purification Technology of Total Flavonoids from Sparganium stoloni by Box-Behnken Design- response Surface Methodology / 中国药房

OBJECTIVE： To optimize the purification technology of total flavonoids from Sparganium stoloniferum. METHODS： Separation and purification by macroporus adsorption resin， using sample solution pH， flow rate and concentration of eluent， the purification rate of total flavonoids as evalution indexes， the purification technology of total flavonoids from S. stoloniferum were optimized by Box-Behnken design-response surface methodology based on single factor test. Validation test was conducted. RESULTS： The optimal purification technology was sample solution pH 4.8， flow rate of eluent 2.0 BV/h， concentration of eluent 72％. The purification rate of total flavonoids in 3 batches of samples was 72.34％ （RSD＝1.77％， n＝3） in validation test， relative errors of which to predicted value （73.99％） was 2.13％. CONCLUSIONS： The optimal purification technology is stable and feasible， and can be used for the purification of total flavonoids from S. stoloniferum.

Decreased gastric body mucosa obestatin expression in abdominal obesity patients with normal body mass index / 生物医学与环境科学（英文）

The aim of the study was to investigate whether the expression of obestatin in gastric body mucosa in abdominal obesity patients with normal body mass index (BMI) is different compared with healthy controls. Twenty abdominal obesity patients with normal BMI and twenty healthy controls were included in the study. The number of obestatin-positive cells in gastric body mucosa was significantly lower in abdominal obesity patients with normal BMI than that in healthy subjects. There was a positive correlation between the numbers of obestatin-positive cells in the gastric body mucosa and plasma obestatin levels in abdominal obesity subjects and control group.

Changes of mobility of kidney tubular epithelial cells in the course of epithelial-mesenchymal transition / 中华实用儿科临床杂志

Objective To evaluate the dynamic changes of cell mobility of renal tubular epithelial cells in the course of epithelial-mesenchymal transition(EMT) and their effect on cell cycle.Methods NRK-52E cells were cultured in vitro and treated with 5 μg/L transforming growth factor(TGF)-β1 to induce EMT.The cell mobility was assessed by using Transwell chamber assay and flow cytometry (FCW) after being treated with TGF-β1 for 4 h,8 h,12 h,24 h and 48 h.The proliferative cell cycle of NRK-52E cells were evaluated by using the FCW.Results 1.EMT was successfully induced by TGF-β1.After being treated by TGF-β1 (5 μg/L),the morphological changes of NRK-52E cells were found with loose cell arrangement and elongated fusiform change in cells body.Meanwhile,after getting treated by TGF-β1,the expressions of E-cadherin protein(epithelial marker) of NRK-52E cells were significantly decreased with time-dependent (P ＜ 0.05),while the expressions of α-smooth musle actin (α-SMA) (mesenchymal cell marker)were significantly increased with time-dependent (P ＜ 0.05).2.The Transwell chamber assay showed that compared with the control group,the cell mobility in the group treated with TGF-β1 was significantly enhanced from 12 h after getting treated with TGF-β1 (P ＜ 0.01).3.The proliferative cell cycle of NRK-52E cells showed no significant difference after being treated with TGF-β1 (P ＞ 0.05).Conclusions The migration ability of the NRK-52E cells are increased incessantly in the course of EMT,which is induced by TGF-β1 without the influence of cell proliferation in vitro.

Interleukin-32 expression in serum of patients with HBV-related liver failure and its significance / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the expression level of IL-32 in serum and its correlation with serum biochemical indices of liver function test and HBV DNA load in patients with HBV-related liver failure.</p><p><b>METHODS</b>Fifty-five patients with HBV-related liver failure (severe hepatitis group) and twenty normal cases (control group) were enrolled in the study. Total RNA in PBMCs was extracted by using TRIzol. IL-32 mRNA level was assayed by using Real-time PCR. IL-32 protein level in serum was detected by ELSIA method. The correlation between IL-32 and ALT, AST, TBIL, HBV DNA load was analyzed using pearson's correlation analysis, respectively.</p><p><b>RESULTS</b>Serum IL-32 expression level in severe hepatitis group was higher than that of control group. Moreover, the difference between them was statistically significant (P < 0.05). Serum IL-32 level was positively correlated with serum ALT, AST, TBIL, respectively (P < 0.05), but was not correlated with HBV DNA load (P > 0.05).</p><p><b>CONCLUSION</b>Serum IL-32 expression level was increased in patients with HBV-related liver failure and was associated with the severity of inflammation. We, therefore, believe that IL-32 might be involved in the pathogenesis of HBV-related liver failure.</p>

Hepatitis B virus X promotes HepG2 cell cycle progression and growth via downregulation expression of p16 protein / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects and related mechanisms of hepatitis B virus X (HBx) protein on cell cycle and growth in hepatocellular carcinoma.</p><p><b>METHODS</b>A human hepatocyte HepG2 cell line stably expressing a green fluorescent protein (GFP)-tagged HBx (HepG2/GFP-HBx cells) was used for the experiment, and HepG2 parental and HepG2/GFP cells was used as the controls. Effect of HBx on cell growth was evaluated by the MTT cell proliferation assay and on cell cycle progression by flow cytometry analysis of cells with or without treatment with 5-aza-2'-deoxycytidine (5-Aza-CdR; 5 pmol/L). Effect of HBx expression on promoter methylation status of the p16INK4A tumor-suppressor gene was detected by methylation-specific polymerase chain reaction and on p16 protein level was analyzed with western blotting.</p><p><b>RESULTS</b>The HepG2/GFP-HBx cells showed significantly higher cell proliferation at 72 hrs of culture (3.225+/-0.038 A490) than either control (HepG2: 2.012+/-0.022 A490, t = -46.86, P less than 0.001; HepG2/GFP: 2.038+/-0.029 A490, t = 42.51, P less than 0.001). The HepG2/GFP-HBx cells also showed significantly lower proportion of cells in the G0/G1 phase (16.45%+/-0.45%) than either control (HepG2: 44.81%+/-1.36%, t = -34.202, P less than 0.001; HepG2/GFP: 42.76%+/-1.58%, t = -28.88, P less than 0.001). However, 5-Aza-CdR treatment did lead to a significant amount of HepG2/GFP-HBx cells being arrested in the G0/G1 phase (33.25%+/-0.79%, t = 31.85, P less than 0.001). The p16INK4A promoter was methylated in the HepG2/GFP-HBx cells, and became demethylation after treatment with 5-Aza-CdR. However, no methylation of p16INK4A promoter was observed in both HepG2 and HepG2/GFP cells. The p16 protein level was significantly lower in the HepG2/GFP-HBx (vs. HepG2 and HepG2/GFP cells) and this level increased after treatment with 5-Aza-CdR.</p><p><b>CONCLUSION</b>HBx protein promotes hepatocellular carcinoma cell cycle progression and growth by shortening the G0/G1 phase, and the underlying mechanism may involve inducing p16INK4A promoter methylation and downregulating p16 protein expression.</p>

Small interfering RNA targeting to hepatitis B virus X gene and 5-aza-2'-deoxycytidineon inhibited growth of the subcutaneous implanted tumor of hepatocellular carcinoma in nude mice / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor effect of small interfering RNA targeting to HBV X gene (X-siRNA) and 5-aza-2'-deoxycytidine (5-aza-dC) on HBV-related hepatocellular carcinoma.</p><p><b>METHODS</b>X-siRNA and control siRNA were synthesized. HepG2/GFP-HBx cells were treated with X-siRNA, and the levels of HBV X mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Nude mice were inoculated with HepG2/GFP and HepG2/GFP-HBx cells subcutaneous respectively to establish implant models of hepatocellular carcinoma, and were treated with X-siRNA, 5-aza-dC alone or in combination, and tumor growth was observed. The methylation of p16 gene promoter was detected by methylation specific polymerase chain reaction (MSP).</p><p><b>RESULTS</b>RT-PCR showed the expression of HBV X mRNA in HepG2/GFP-HBx cells was inhibited markedly by X-siRNA. The nude mice experiment showed that the gross tumor volume was much bigger in HepG2/GFP-HBx group than that in HepG2/GFP group (P < 0.05). The growth of palpable tumors in X-siRNA or 5-aza-dC treatment group notably decreased (P < 0.05). MSP analysis showed that p16 gene methylation was observed in HepG2/ GFP-HBx-caused palpable tumors, while no methylation was detected in HepG2/GFP group. However, after treatment with X-siRNA or 5-aza-dC, p16 gene methylation reduced.</p><p><b>CONCLUSIONS</b>HBV X-siRNA and methylation inhibitor can inhibit the growth of hepatoma cells via reversing p16 methylation.</p>

Expression and biological activity identification of recombinant Hap protein of NTHi / 南方医科大学学报

<p><b>OBJECTIVE</b>To express and purify Hap protein of nontypeable Haemophilus influenzae (NTHi) in prokaryotic system, and study its immunogenicity and adhesive activity.</p><p><b>METHOD</b>Hap protein was expressed in E.coli BL21 with pET32a (+)-Hap and purified by affinity chromatography. The adhesive activity of the recombinant Hap protein was observed in competitive adhesion assay using scanning electron microscope and bacterial counting. BALB/C mice were immunized intranasally with the purified recombinant Hap protein and cholera toxin B subunit (CT-B), and anti-Hap IgA and IgG were detected by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>SDS-PAGE analysis showed a single band of the target protein, whose purity reached 85% according to the result of Gel analysis software. The concentration of the protein was 3.2 g/L after ultrafiltration and condensation. Competitive adhesion assay showed that compared with control group, the recombinant Hap protein significantly inhibited the adhesion of NTHi to ECM (P<0.01). Compared with Hap immunization alone, immunization with Hap combined with CT-B resulted in significantly higher titers of anti-Hap IgG and IgA in mice (P<0.05).</p><p><b>CONCLUSION</b>Highly purified recombinant Hap protein has been obtained in a prokaryotic system and shows good immunogenic and adhesive activities. These results will establish the basis for further study of NTHi vaccine.</p>

Cloning Hap gene from non-typeable Haemophilus influenzae and expression of Hap protein in prokaryotic cell / 生物医学工程学杂志

This study was aimed to construct a prokaryotic expressing vector of Hap gene from Nontypeable Haemophilus influenzae, and express and identify the fusion proteins of Hap-His in E. coli. The gene encoding protein Hap was amplified from Nontypeable Haemophilus influenzae ATCC49247 chromosomal DNA by PCR, then it was cloned into prokaryotic expression plasmid pET-32a (+). The recombinant plasmid pET-32a(+)-Hap was transformed into E. coli BL21 and expression was induced by Isopropy-beta-D-thiogalatoside(IPTG). The Hap-His fusion protein expressed so was analyzed by SDS-PAGE and Western-blot. The results showed that the recombinant expressive plasmid pET-32a (+)-Hap was constructed successfully, and the recombinant plasmid expressed Hap-His fusion protein with relative molecule mass 176 000 and mainly existed in inclusion body. This fusion protein could combine with anti-His monoclonal antibody specifically through Western blot analysis. Successful expression of Hap-His fusion protein in prokaryotic cell could lay a basis for further study of immunocompetence of Hap protein and development of NTHi vaccine.

Observation on therapeutic effect of eye-needling combined with medication for treatment of ophthalmoplegia / 中国针灸

<p><b>OBJECTIVE</b>To observe the therapeutic effect of eye-needling combined with medication for treatment of ophthalmoplegia and explore the possible mechanism.</p><p><b>METHODS</b>One hundred and twenty cases were randomly divided into a treatment group and a control group. According to etiological factors, the control group were treated with medication and the treatment group with the medication plus eye-acupuncture at main point ocular muscles. Changes of the rima oculi, the range of ocular movement and the dialopia angle after treatment were recorded and statistically analyzed in the two groups.</p><p><b>RESULTS</b>The total effective rate was 93.4% and the cured rate was 54.1% in the treatment group, and 74.6% and 18.6% in the control group, with significant difference between the two groups (P < 0. 01).</p><p><b>CONCLUSION</b>Eye-needling combined with medication has an obvious therapeutic effect which is better than simple medication for ophthalmoplegia.</p>

Preparation and characterization of a monoclonal antibody against human SOCS3 / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare and characterize the monoclonal antibody (mAb) against human SOCS3.</p><p><b>METHODS</b>BALB/c mice were immunized with recombinant GST-SOCS3 protein, from which the spleen cells were isolated and fused with Sp2/0 cells. After several rounds of screening and cloning, the hybridoma cell strain secreting anti-SOCS3 mAb was obtained, whose specificity was evaluated using ELISA and Western blotting, and the titer, immunoglobulin subtype and affinity of the mAb were also measured.</p><p><b>RESULTS</b>The hybridoma cell strain secreting anti-SOCS3 mAb was identified to belong to IgG2a subtype. The mAb titers in cultural supernatant and acetic fluid were 1:640 and 1:25600, respectively, as determined by ELISA with affinity reaching 4.84x10(6) L/mol.</p><p><b>CONCLUSION</b>The success in anti-SOCS3 mAb preparation provides the basis for further study of the negative regulation of cytokine signal transduction and the immunoregulation in microorganism infections.</p>

Optimized isolation and purification of non-typeable Haemophilus influenzae Haps protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To optimize the isolation and purification conditions for Hap(s) protein of non-typeable Haemophilus influenzae.</p><p><b>METHODS</b>Hap(s) protein was purified by ammonium sulfate precipitation, dialysis desalting and Hitrap weak cation exchange columns of CM Sepharose Fast Flow. The condition of the elution was optimized for pH and ionic strength, the absorbance at 280 nm of the elution samples were detected, and the targeted protein band in the collected samples was observed by SDS-PAGE electrophoresis.</p><p><b>RESULTS</b>The Hitrap ion exchange column was eluted with buffer 1, which resulted in a baseline distribution of absorbance at 280 nm. Buffer 2 elution of the column resulted in the presence of peak absorbance with trails, which was identified to be constituted by some low molecular weight bands by subsequent SDS-PAGE. In serial column elution with buffer 3 with different ionic strength, a peak absorbance was observed with the ionic strength of 100 mmol/L NaCl, and SDS-PAGE confirmed that the peak was generated by the target protein. No obvious peaks or bands in SDS-PAGE occurred with the other ionic strengths.</p><p><b>CONCLUSION</b>The pH of the buffer only affect the elution of the irrelevant proteins rather than the Hap(s) protein, and elution with the buffer containing 100 mmol/L NaCl can be optimal for eluting the Hap(s) protein.</p>

Oxidative damage to DNA and its relationship with diabetic complications / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To detect the oxidative DNA damage in diabetic patients and to investigate the relationship of oxidative DNA damage with diabetes and diabetic nephropathy.</p><p><b>METHODS</b>Single cell gel electrophoresis (SCGE) was used to detect the DNA strand breaks in peripheral blood lymphocytes, and oxidative DNA damage product and serum 8-OHdG were determined by a competitive ELISA in 47 cases, including 25 patients without diabetic complications, 22 patients with diabetic nephropathy and 25 normal control subjects.</p><p><b>RESULTS</b>Diabetic patients showed greater oxidative damage to DNA. The percentage of comet cells and the length of DNA migration (comet tail length) of peripheral blood lymphocytes were significantly increased in patients with diabetes, and significantly higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05). There was a significant increase in serum 8-OHdG in diabetic patients compared with normal subjects (P < 0.05). Moreover, serum 8-OHdG was much higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05).</p><p><b>CONCLUSION</b>There is severe oxidative DNA damage in diabetic patients. Enhanced oxidative stress may be associated with diabetes, especially in patients with diabetic nephropathy.</p>