Correlation between Serum Osteoprotegerin/Soluble Receptor Activator of Nuclear Factor-κβ Ligand Levels and the Left Ventricular Diastolic Dysfunction in Type 2 Diabetes / 中山大学学报(医学科学版)

ObjectiveTo study the possible correlation between serum osteoprotegerin （OPG）/soluble receptor activator of the nuclear factor κB ligand （sRANKL） levels and the left ventricular diastolic dysfunction （LADD） in patients with type 2 diabetes mellitus （T2DM）. MethodsTotally 68 T2DM patients and 37 healthy controls were selected. Serum OPG and sRANKL were determined by solid-phase enzyme-linked immunosorbent assay （ELISA）. The left ventricular diastolic function of T2DM patients was measured by transthoracic echocardiography， where E/A < 1 were regarded as LVDD. T2DM patients were further divided into two subgroups according to E/A ratio （E/A≥1.0 and E/A<1）. Spearman correlation analysis， logistic regression and ROC curves were used to assess the possible correlation between serum OPG/sRANKL and LADD in T2DM patients. ResultsCompared with the healthy controls， serum OPG level in T2DM patients was higher with statistically significant difference （P <0.01）， while serum sRANKL level was lower without statistically significant difference （P =0.32）. T2DM patients with E/A<1 had significantly higher OPG level and lower sRANKL level than those with E/A≥1（P <0.01） in subgroup analysis. Spearman correlation analysis showed serum OPG level was negatively correlated with E/A ratio， while sRANKL was positively related with E/A ratio. In single factor logistic regression analyses， serum OPG ［OR （95% CI）=1.068 （1.031， 1.106）， P<0.001］ and sRANKL ［OR （95% CI）=0.976 （0.959， 0.992）， P=0.003］ were significant correlation with LVDD in T2DM patients. ROC curve analysis showed that the sensitivity and specificity of combined OPG and sRANKL in diagnosing T2DM patients LADD were 78.13% and 88.3%， respectively （area under the curve： 0.857； 95% CI=（0.768， 0.946）； P<0.001）. ConclusionsThe elevated OPG and decreased sRANKL levels may be associated with LADD in T2DM patients.

Gene Analysis for the Sebaceous Carcinoma of Scalp by Whole Exome Sequencing / 中山大学学报(医学科学版)

ObjectiveTo reveal the differences of the related pathogenicity gene mutations between sebaceous adenocarcinoma （SC） of scalp and sebaceous adenoma （SA） of scalp on whole exome level. MethodsWhole exome sequencing was performed on a SC sample and a SA sample by Illumina Hiseq 2500 platform. Suspicious single nucleotide variation sites were selected for mutation conservation and functional analysis. SciClone was used to track subclone evolution and clonal map information was obtained for each tumor sample. The high-frequency significant gene mutations in the tumor sample were screened by MutSigCV software， and compared with the known driver genes. ResultsTwo driver genes TFDP1 and ACVR1B harboring mutations in scalp SC compared to SA were found. ConclusionsThe finding of mutation in driver genes TFDP1 and ACVR1B should be confirmed in a large cohort， which might reveal the mechanism of scalp SC development and find a therapeutic target for SC.

Effect of chronic exposure to ultraviolet on skin barrier function / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of chronic ultraviolet (UV) exposure on skin barrier function and photoaging process.</p><p><b>METHODS</b>One hundred and fifty-six volunteers from Hanghzou areas were enrolled in the study. UV-exposed skin areas (neck, dorsum of hand or frontier chest) and UV-unexposed areas (waist, buttock or abdomen) were tested. Probe CM 825 of skin multi-functional detector MPA9 was applied to test the skin water content; probe TM 300 was applied to test transepidermal water loss (TEWL) and probe RVM 600 was applied to detect skin elasticity (Ur/Uf). Relative perfusion unit (PU) of the skin was detected by laser doppler flowmetry (LDF).</p><p><b>RESULT</b>Skin water content value at UV-exposed skin areas was 12.78 ± 2.36 in elderly group (>50y), which was significantly lower than that of UV-unexposed skin areas(23.68 ± 3.24, P= 0.036). Highest level of TEWL (12.98 ± 2.86) g . m(-2) . h(-1) was detected at UV-exposed areas in elderly group; there were trends of increasing TEWL levels at UV-exposed areas than at UV-unexposed areas in all age groups, however, there were no statistical differences (P>0.05). The elasticity of Ur/Uf value at UV-exposed skin areas in elderly group was 0.11 ± 0.07, which was remarkably lower than that of UV-unexposed skin areas (0.32 ± 0.1, P=0.028). No significant difference of skin perfusion was observed between UV-exposed and UV-unexposed areas.</p><p><b>CONCLUSION</b>Chronic exposure to UV may damage skin barrier function and therefore play a role in skin photoaging process.</p>

Telomerase expression is not involved in aging process of human keratinocytes induced by UVB irradiation / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the mechanism involved in aging process of immortalized human keratinocyte (HaCaT) and primary human epidermis keratinocyte of adults (HEKa) irradiated by ultraviolet B(UVB).</p><p><b>METHODS</b>HEKa and HaCaT were repeatedly exposed to UVB at a subcytotoxic level. SA-beta-Gal staining was performed to evaluate the senescence state; flow cytometry was applied to detect the changes of apoptosis, necrosis and cell cycle. Intracellular levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by ELISA method. Western blot was performed to detect the expression pattern of redox protein p66Shc and RT-PCR was performed to determine the mRNA level of human telomerase reverse transcriptase (hTERT).</p><p><b>RESULT</b>Strong positive SA-beta-Gal staining was observed in both HEKa cell and HaCaT cells after UVB irradiation. Apoptosis rate increased from (1.81 +/-0.25)% to (4.43 +/-0.28)% and necrosis rate increased from (0.05 +/-0.01)% to (0.10 +/-0.03)% in HaCaT cell, but no marked arrest of cell cycle was observed during UVB irradiation. As a contrast, apoptosis rate of in HEKa cells significantly increased from (0.65 +/-0.05)% to (59.53 +/-2.35)%, and the necrosis rate in HEKa cells also reached (3.89 +/-0.24)%(P<0.05). Growth arrest in G0/G1 phase was also found in HEKa cells. In both cell lines, intracellular level of SOD decreased and MDA increased remarkably after UVB exposure, and an increased expression of p66Shc protein was also observed. High level of hTERT mRNA was detected in HaCaT cells and UVB exposure had little effect on its expression.</p><p><b>CONCLUSION</b>The stress-induced premature senescence (SIPS) in HaCaT and HEKa cell lines by UVB irradiation might be closely associated with increased intracellular levels of oxidative stress, not related to the telomerase expression.</p>

Survey on skin aging status and related influential factors in Southeast China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To investigate cutaneous aging patterns of residents in Hangzhou, Zhejiang, China, and their contributing factors.</p><p><b>METHODS</b>Eight hundred and forty-eight Hangzhou residents received the survey between March 2004 and September 2004.</p><p><b>RESULTS</b>Facial wrinkling first occurred at 21 years of age and skin elasticity began to lose at 22 years of age. In middle-aged and old people, facial wrinkling and looseness escalated with the increase of ultraviolet (UV)-exposure time, indicating the accelerating effect of a higher accumulative dose of UV radiation on skin aging. Only Fitzpatrick types II, III and IV were found in the skin phototypes of residents in Hangzhou area, and Fitzpatrick type II seemed to be much more subject to severe wrinkling, elasticity destruction and skin tumors than types III and IV. The oily skin was more protected against wrinkling and facial looseness than dry skin. However, as to concomitant cutaneous diseases, no difference was found among different skin types.</p><p><b>CONCLUSION</b>Age, solar-exposure time, Fitzpatrick type and skin type are the associated forces in promoting skin aging, and emotional factor seems to be another independent risk factor. The age of 49 years and 2 h/d of solar-exposure time seem to be the turning points responsible for dramatic changes of cutaneous appearance in the process of skin aging in Southeast China.</p>

Evaluation on the accuracy of obese indexes in predicting obstructive sleep apnea-hypopnea syndrome in male adults / 中华流行病学杂志

<p><b>OBJECTIVE</b>To determine the best cutoff value and reference standard of obese indexes and to compare their diagnostic value in screening, predicting and diagnosing obstructive sleep apnea-hypopnea syndrome (OSAHS) in male adults by receiver operating characteristic curve (ROC).</p><p><b>METHODS</b>A total of 1110 male adults were included and cross-sectional study was adopted. Four parameters including: body mass index (BMI), neck circumference (NC), waist circumference (WC) and waist-to-hip ratio (WHR) were studied. The correlativity between apnea-hypopnea index (AHI) and BMI, NC, WC and WHR were analyzed by pearson correlation test. ROC curve was used to assess the diagnostic value and determine their best cutoff value to predict OSAHS and reference standard to screen/diagnose it. Their conformity in predicting OSAHS was analyzed by area under curve (AUC). SPSS software version 13.0 was used for statistical analysis and P< or =0.05 was considered to indicate statistical significance.</p><p><b>RESULTS</b>(1) All of the fourindexes (BMI, NC, WC and WHR) showed a significantly positive correlation with AHI in adult male patients, with their Pearson coefficients as 0.373, 0.276, 0.291 and 0.127, respectively, and their P value were all below 0.001; (2) The best cutoff value of BMI, NC and WC in predicting OSAHS were 26.5 kg/m2 , 39 cm, 95 cm, and their corresponding sensitivity rates were 54%, 57% and 64% while rates of specificity as 73%, 65% and 53%; (3) BMI was better than other two indexes in predicting OASHS in adult male patients; (4) The value of BMI, NC and WC in screening OSAHS among adult male patients with 90% sensitivity and 10% rate of missed diagnosis as 23 kg/m2, 35 cm, 85 cm; (5) The value of BMI, NC and WC in screening OSAHS in adult male patients with 90% specificity and 10% misdiagnosis rate as 29 kg/m2, 43 cm, 105 cm, respectively.</p><p><b>CONCLUSION</b>BMI seemed better in predicting OSAHS in male adults than NC, WC and WHR. The best cutoff values to predict OSAHS of BMI, NC and WC were 26.5 kg/m2, 39 cm and 95 cm, respectively. Their reference standards to screen OSAHS were 23 kg/m2, 35 cm and 85 cm while 29 kg/m2, 43 cm and 105 cm to predict OSAHS.</p>

Effects of angiotensin II on connexin 43 of VSMCs in arteriosclerosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To observe the effect of angiotensin II (Ang II) on expression of gap junction channel protein connexin 43 (Cx43) in the proliferation process of vascular smooth muscle cells (VSMCs) during the early stage of arteriosclerosis.</p><p><b>METHODS</b>Thirty-two adult male rabbits were randomly divided into 4 groups. Rabbits in Group A were fed common diet while others in Groups B, C, and D were fed high-cholesterol diet. Losartan (10 mg/(kg.d)) and ramipril (0.5 mg/(kg.d)) were added in the diet of Groups C and D, respectively. The animals were sacrificed after 8 weeks and abdominal aortas were removed and dissected. The expression of Cx43 was assayed using RT-PCR and Western Blotting analysis.</p><p><b>RESULTS</b>Cx43 was increased markedly in both protein and mRNA level in Groups B, C, and D fed high-cholesterol diet compared with that in control group (P<0.01). Cx43 level in losartan or ramipril treated groups was higher than that in control group (P<0.01, P<0.05), but lower than that in high-cholesterol diet groups (P<0.05, P<0.01).</p><p><b>CONCLUSION</b>Cx43 level was upregulated in VSMCs during early atherosclerosis. Losartan and ramipril can inhibit the expression of Cx43.</p>

Inhibitory effects of mizolastine on substance P-induced production of leukotriene B4 and interleukin 5 in mouse skin / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To observe the inhibitory effect of mizolastine on substance P(SP)-induced production of leukotriene B(4) (LTB(4)) and interleukin 5 (IL-5) in mouse skin.</p><p><b>METHODS</b>Mice were fed with different doses of mizolastine or other control drugs, 30 min after administration animals were injected intradermally with SP on the back. The treated skin samples were taken and competitive enzyme-link immunoassay (ELISA) method was applied to detect LTB (4) and IL-5 in the skin samples.</p><p><b>RESULT</b>The LTB(4) and IL-5 levels in 10 mg/kg mizolastine group were (1.23 +/-0.29)pg/ml and (34.28 +/-11.00)pg/ml, respectively, which were lower than those in saline control group [(5.52+/-1.88)pg/ml and (179.62 +/-46.25)pg/ml respectively] or loratadine group [(3.89+/-1.27)pg/ml and (127.74 +/-43.27)pg/ml respectively]. No significant difference was found between 10 mg/kg mizolastine group and dexamethasone group (P=0.161 and P=0.508).</p><p><b>CONCLUSION</b>Mizolastine might inhibit the production of LTB(4) and IL-5 induced by substance P in mouse skin, suggesting that anti-inflammatory effect and the blockade of histamine H1 receptors might be involved in its anti-pruritic mechanisms.</p>

Effects of fructose-1,6-diphosphate on concentration of calcium and activities of sarcoplosnic Ca2+-ATPase in cardiomyocytes of Adriamycin-treated rats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To observe the effects of fructose-1,6-diphosphate (FDP) on serum levels of cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB), as well as the concentration of calcium in cardiomyocytes (Myo[Ca(2+)]) and activity of sarcoplosnic Ca(2+)-ATPase (SRCa(2+)-ATPase) in Adriamycin (ADR)-treated rats.</p><p><b>METHODS</b>Rats were intraperitoneally injected with ADR (2.5 mg/kg every other day for 6 times) and then with different dosages of FDP (every other day for twenty-one times). Bi-antibodies sandwich Enzyme linked immune absorption assay (ELISA) was performed to detect serum level of cTnI. CK-MB was detected by monoclonal antibody, Myo[Ca(2+)] was detected by fluorescent spectrophotometry and the activity of SRCa(2+)-ATPase was detected by inorganic phosphate method.</p><p><b>RESULTS</b>FDP (300, 600, 1200 mg/kg) significantly reduced the serum levels of cTnI and CK-MB, while at the same time decreased calcium concentration and increased SRCa(2+)-ATPase activity in cardiomyocytes of ADR-treated rats (P<0.01).</p><p><b>CONCLUSIONS</b>FDP might alleviate the cardiotoxic effects induced by ADR through decreasing calcium level as well as increasing SRCa(2+)-ATPase activity in cardiomyocytes.</p>

Effects of nasal continuous positive airway pressure short-term treatment on C-reactive protein and intercellular adhesion molecule-1 in patients with obstructive sleep apnea-hypopnea syndrome / 中南大学学报(医学版)

OBJECTIVE@#To investgate the effects of nasal continuous positive airway pressure (nC-PAP) short-term treatment on the serum level of C-reactive protein (CRP) and intercellular adhesion molecule-1 in obstructive sleep apnea-hypopnea syndrome (OSAHS) patients.@*METHODS@#Twenty patients with OSAHS ( AHI > or = 15 ) and 15 controls (AHI < 15) were recruited. After polysomnography (PSG), the venous blood was collected from all subjects to investigate CRP and ICAM-1. The effects of short-term treatment (4 d) of nCPAP on the serum levels of CRP and ICAM-1 were studied in patients with moderate and severe OSAHS.@*RESULTS@#Before the treatment, there was a significant positive correlation between CRP and AHI in all subjects (r = 0.615, P < 0.001), a significant negative correlation between CRP and the mean nocturnal oxygen saturation (r = -0.682, P < 0.001), and a significant negative correlation between CRP and the lowest nocturnal SaO2 (r = -0.61, P < 0.001). There was a significant positive correlation between ICAM-1 and AHI in all subjects (r = 0.751, P < 0.001), a significant negative correlation between ICAM-1 and the mean nocturnal oxygen saturation (r = -0.68, P < 0.001), and a significant negative correlation between ICAM-1 and the lowest nocturnal oxygen saturation (r = -0.656, P < 0.001). There was a significant positive correlation between CRP and ICAM-1 (r = 0.437, P = 0.009). The levels of CRP and ICAM-1 were significantly higher in patients with moderate and severe OSAHS than in the controls (P < 0.01). nCPAP decreased the levels of CRP and ICAM-1 in patients with moderate and severe OSAHS (P < 0.01).@*CONCLUSION@#Inflammatory reaction exists in OSAHS and can be palliated after nCPAP short-term treatment.

Construction of small interfering RNA targeting heparanase gene and its inhibitory effect on the in-vasiveness of human malignant melanoma cell line A375 in vitro / 中华皮肤科杂志

Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P

Expression of B7-CD28/CTLA-4 in peripheral blood lymphocytes of female patients with genital condyloma acuminatum / 中华皮肤科杂志

Objective To investigate the significance of B7-CD28/CTLA-4 (CD152),co-stimulatory molecules of T lymphocytes,in the onset,development and prognosis of genital congdyloma acuminatum (CA) in females.Methods Flow cytometry was utilized to detect the expression levels of CD80/CD86 in peripheral blood lymphocytes and of CD28/CD152 (CTLA-4) in CD4~+/CD8~+ T lymphocytes from 30 CA patients (17 primary CA,13 recurrent CA,15 at recovery stage of CA) and 15 healthy volunteers as con- trols.Results No significant difference was found for the frequencies of CD80~+,CD86~+,CD4~+/CD28~+ and CD8~+/CD28~+ lymphocytes between the primary or recurrent group and the control group.The frequencies of CD8~+/CD28~+,CD4~+/CD28~+ and CD80~+ lymphocytes were significantly higher in the recovery group than those in the recurrent group (P0.05).Conclusions There is an abnormal expression of co-stimulatory molecules B7-CD28/CTLA-4 (CD152) in periphoral blood lym- phocytes,CD4~+ and CD8~+ T lymphocytes in female patients with genital CA,and the expression abnormaility is closely linked with different disease stages of CA.

The Progress of Recombinant Escherichia coli for Production of Succinic Acid / 微生物学通报

Succinic acid is regarded as C4 platform chemical, which has a huge potential market with wide applications in biopolymer, food, medicine, and so on. Production of succinic acid by microorganism fer-mentation has attracted much attention in recent times. By fermentation, succinic acid could be produced from renewable resources in place of petroleum with low cost and low pollution; especially CO2 could be fixed by microorganism to form the product, which is a promising way to deal with the green gas. Some E. coli have been chosen as good succinic acid producing strains because of their clear genetic backgrounds, convenience to be modified and controlled, and good growth properties with low nutrients requirements. The progress of genetic engineering strategy and metabolic engineering technology for construction of succinic aicd producing recombinant E. coli has been introduced in this paper with the discussion of the future re-search in this area.