Objective To explore the correlation between plasma Klotho and FGF23 levels with senile hypertension .Methods Totally 180 cases of hypertension aged more than 60 years old were selected as the research group and divided into the grade 1 hypertension group(60 cases) ,grade 2 hypertension group(60 cases) and grade 3 hypertension group(60 cases) .The age and pro‐portion of male to female were basically consistent .60 elderly people without hypertension were selected as the control group .The level of plasma Klotho and FGF23 protein were tested by ELISA .The lipid level was detected by the automatic biochemical analy‐zer .The detection results were compared with those in the control group for analyzing their correlation .Results Plasma Klotho protein level in each hypertension group was lower than that in the control group ,while plasma FGF23 protein level and BMI were significantly higher than those in the control group(P<0 .05) .With the blood pressure level increase ,plasma Klotho protein level was gradually decreased ,while FGF23 protein level was gradually increased .The negative correlation was found between plasma Klotho protein and FGF23 protein (r= -0 .282 ,P=0 .001) .Plasma Klotho protein and FGF23 protein had no obvious correlation with BMI (r= -0 .063 ,-0 .098 ,P>0 .05) .Conclusion Plasma Klotho protein and FGF23 protein levels have obvious correlation with the blood pressure level ,which may jointly participate in the occurrence and development of hypertension .

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective:To evaluate the application value of robost optimization of brachytherapy for cervical cancer.Methods:Twenty patients who completed radical treatment were recruited in this study. The dose volume histogram (DVH) parameters were statistically compared between the conventional and robust optimization plans, and the robustness between the conventional and robust optimization plans was evaluated using DVH and DVH bands. The robust optimization method utilized the worst dose distribution to consider the dose in the presence of uncertainties. In each optimization iteration, the dose distributin when the radioactive source shifted along the X, Y, and Z directions (±2 mm), and the dose distribution when the radioactive source was not shifted were calculated. The worst dose distribution for each voxel was the lowest dose in the target and the highest dose outside the target under all circumstances. The iterative objective function was calculated by the worst dose distribution.Results:In the scenario of no shifting of radioactive source position, the mean value of robust optimization was significantly lower and that of V 150% was significantly higher than those of conventional optimization (both P<0.05). When considering the shifting of radioactive source position, the worst dosimetric parameters of multiple dose distributions were statistically compared. The mean HR-CTV D 100% values did not significantly differ between the robust and conventional optimization plans, whereas the mean D 90% value (range: 0.02-0.03 Gy) of robust optimization was significantly higher than that of conventional optimization ( P<0.05). Robust optimization increased the D 2cm 3 of the bladder and small intestine, and the rectum dose was increased with the shifting of the radioactive source position in the robust optimization. The DVH bands did not significantly differ between the conventional and robust optimization plans for all patients. Conclusions:Robust optimization based on the worst dose distribution fails to significantly improve the robustness of brachytherapy for cervical cancer. Alternative methods are required to minimize the dosimetric effect of uncertainties in brachytherapy.

OBJECTIVEThis paper is to explore the relationships between the occurrence of structural birth defects and environmental risk factors on birth defects.

METHODSA survey was conducted by using a population-based case control study. A total number of 388 structural birth defects, which were detected after 28 weeks' gestational age to 7th day after birth in various hospitals were recruited in Wuxi from 2002 to 2004. The control group consisted of 1607 normal live births born in the same period. Multiple logistic regression model was used to analyze relationships between environmental risk factors and occurrence of structural birth defects after controlling women's education level and childbearing age.

RESULTSAmong those factors which affected birth defects, the exposure to toxic substances in working environment was the highest risk factor, with OR value as 5.37 (95 % CI : 3.60-7.99). Suffering from reproductive tract infections, high fever during first trimester gestation, and taking potential teratogenic drugs such as hypnotics and anti-convulsion drug agents, might significantly increase the risk of structural birth defects, with OR values (95% CI) as 3.38 (1.33-8.56), 3.57 (1.73-7.37) and 2.75 (1.57-4.83) respectively. However, the correlation between oral contraceptives intake within six months before pregnancy and risk of birth defects had no statistical significance. In addition, pollution of the residential environment, staying up all night and raising pets at home, had relatively lower association with birth defects, with OR values (95% CI) as 2.28 (1.75-2.98), 1.96 (1.21-3.18) and 2.50 (1.66-3.76) respectively, yet with a higher proportion of exposure to those factors in pregnant women. There was a significant increase in occurrence risks of birth defects with increase in the number of exposure to environmental risk factors. OR values for having one, two, three kinds of environmental risk factors were 1.60, 4.32 and 10.23 respectively.

CONCLUSIONThe common structural birth defects were affected by a number of environmental risk factors. There was also a dose-response relationship between the number of environmental risk factors and occurrence risks of structural birth defects noticed in our study.

Air Pollution ; adverse effects ; Case-Control Studies ; Congenital Abnormalities ; etiology ; Female ; Humans ; Logistic Models ; Maternal Exposure ; adverse effects ; Pregnancy ; Risk Assessment ; Risk Factors

Objective:To evaluate the clinical efficiency and prognostic factors of autologous peripheral blood stem cell trans-plantation (APBSCT) in 30 cases of multiple myeloma (MM). Methods:Two of the 30 patients received the second treatment of APB-SCT because of relapse after the first treatment. Thus, a total of 32 case-times of APBSCT were studied. Combination chemotherapy was inducted regularly before APBSCT (11 patients used bortezomib as an induction drug), and chemotherapy combined with the G-CSF regimen was used to mobilize peripheral blood stem cells. Preconditioning was based on melphalan. Results:Mononuclear cells in harvest were 6.41 × 108/kg, and CD34+cells in harvest were 4.75 × 106/kg. The median times of neutrophil and platelet engraftment were 9.5 and 11 d, respectively. The complete remission (CR) and very good partial remission (VGPR) rates were 37.5%and 34.4%af-ter APBSCT, respectively. The median overall survival (OS) was 67.27 months in all patients, and the median progression-free survival (PFS) was 29.77 months. The median PFS rates were 29 and 20 months in the patients who achieved CR and PR, respectively, and the median PFS was not observed in the patients who achieved VGPR. Statistical differences in PFS were detected between the CR+VGPR and PR groups (P=0.025). The CR rates were 63.6%and 23.8%in the bortezomib (bortezomib-based chemotherapy) and non-bortezo-mib groups (P=0.034), respectively. The median OS and PFS were not obtained in the bortezomib group, whereas the median PFS was 22 months in the non-bortezomib group (P=0.045). Conclusion: MM patients treated with bortezomib-based chemotherapy followed by APBSCT had prolonged PFS. APBSCT can improve the response and survival of MM patients.

Objective To provide updated recommendations for further adjustment of iodine deficiency disorders control strategy by evaluating awareness of the dangers of iodine deficiency disorders and their prevention and control measures among local residents and the willingness of choosing iodized or non-iodized salt in urban communities of Jiangsu province.Methods Trained health service workers did questionnaire investigation on iodine deficiency disorders prevention knowledge by calling computer-randomly selected family telephone numbers in every two blocks of provincial capital (Nanjing) and coastal city (Nantong) in Jiangsu province in 2010.The survey topics covered include:when supplied with both non-iodized and iodized salt,what kind of salt the local residents chose and the reasons; residents awareness of the dangers of iodine deficiency disorders and their prevention and control measures; resident's personal information(including only occupational category and age group).Key fields were rechecked from 10％ randomly selected valid questionnaires.Results Four hundred and fifty-five valid questionnaires were obtained by calling 2021 randomized telephone numbers in Nanjing and Nantong cities.After 10.1％(46/455) of the valid questionnaires were rechecked,the total coincidence rate was 87.0％(40/46).In the two cities,there were 73.2％ (333/455) of those 455 respondents saying they would choose iodized salt when iodized and non-iodized salt were supplied at the same time.There were 69.0％(314/455) of respondents had heard of iodine deficiency disorders,of which 79.6％(250/314) of them were aware of the dangers of iodine deficiency is endemic goiter,and 10.8％ (34/314) of them knew that iodine deficiency can cause varying degrees of mental damage.With respect to useful preventive methods of iodine deficiency disorders,there were 69.4％ (218/314) respondents know that eating iodized salt,and 41.1％ (129/314) respondents know that taking laver or kelp would prevent iodine deficiency disorders.Conclusions Remarkable achievements on iodine deficiency disorders health promotion and health education have been gained in urban communities of both the provincial capital and the coastal city Jiangsu province.However,some of the people are still not very clear why we should eat iodized salt.Salt industry in the current management system,is still premature to provide iodized salt and non-iodized salt simultaneously now.

Background The diagnosis and treatment of fungal keratitis are knotty.There is no quantitative method to identify the disease and judge the therapeutic effect of the antifungal agent.Studies have determined that serum (1,3-) β-D-glucan level can sensitively and specifically reflect the state of systemic mycotic-causing diseases.However,whether (1,3-) β-D-glucan level in tear can monitor and diagnose mycotic keratitis is unclear.Objective Purpose of this study was to investigate the change of tear (1,3-) β-D-glucan level following the administration of antifungal drug in fungal keratitis patients,and evaluate the diagnosis and monitor value of (1,3-) β-D-glucan in tears for fungal keratitis.Methods Sixty patients who were diagnosed as fungal keratitis by fungal culture were analyzed in Affiliated Hospital of Qingdao University Medical College from July 2010 to May 2012.The patients received the topical administration of antifungal drug for 28 days.Thirty healthy volunteers without eye disease served as normal controls.The tear of 50 μl was collected from each subject for the detection of (1,3)-β-D-glucan before the therapy,7,14,28 days after therapy and 7 days,14 days after the drugs were stopped,respectively.The dynamic changes of (1,3-) β-D-glucan levels in tears were evaluated and compared with the manifestation of the lesions under the laser scanning confocal microscope.The patients without hyphal by the laser scanning confocal microscopy and tear (1,3-)β-D-glucan level less than 20 ng/L were subsequently treated for another 7 days,and the following-up duration was 2 months.The informed consent was obtained before any medical examination was performed from each subject.Results (1,3-)β-D-glucan level in tears (Log value) was (6.37 ±0.48)ng/L in the patient group,and was significantly higher than (2.00±0.31) ng/L in the normal control group (t =2.89,P＜0.01).The lesion was smaller with the gradually clear border,and the number of mycelia was decreased under the laser scanning confocal microscope 7 days after treatment.(1,3-) β-D-glucan level in tears was gradually declined in a time-dependent manner after treatment.The (1,3)-β-D-glucan level in tears (Log) was (5.19 ± 0.42),(4.16 ± 0.33),(2.99 ±0.42),(2.91 ±0.39),(2.80±0.40) ng/L 7,14,28 days after treatment,and 7 days,14 days after the drugs were stopped,respectively,with a statistically significant difference in comparison with (6.37±0.48)ng/L before treatment (P＜0.01).(1,3)-β-D-gluean level in tears remained a lower level till the end of follow-up,and no recurrence of lesion was found in the patient group.Conclusions Detecting (1,3)-β-D-glucan level in tears is of good diagnosis and monitor value in the evaluation of fungal keratitis.

Objective To investigate the expression of inregrin?_3 and integrin?_1 in breast cancer and its bio- logical significance.Methods Immunohistochemical assay was used to determine the expression of integrin?_3 and integrin?_1 in the breast cancer(32 cases).Results In normal breast tissue,the positive expression rates of integrin?_3 and integrin?_1 were 0 % and 25 %.In the breast cancer tissue,the positive expression rates of integrin?_3 and inte- grin?_1 were 36 % and 81%.Conclusion The integrin?_3 and integrin?_1 are close associated with the biological sig- nificance of breast cancer.To examine its expression is useful to evaluate the aggressive degree,metastatic potential and prognosis in patients with breast cancer.

UNLABELLEDTo identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp.

CONCLUSIONSSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.

Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Fetus ; Gene Library ; Humans ; Mesoderm ; cytology ; metabolism ; Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism

Objective:To investigate the effect of concentrated growth factor (CGF) on proliferation and differentiation in Beagle adipose-derived stem cells (ADSCs).Methods:ADSCs were isolated from adipose tissue of healthy Beagles and cultured.The multidirectional differentiation potential of ADSCs was identified.The ADSCs were assigned to a CGF group and a control group.The rate of proliferation was analyzed by CCK-8 assay.The osteogenic differentiation capability was detected by ALP staining after the osteoinduction.Bone formationrelated gene expression was detected by RT-PCR.Results:CGF promoted the proliferation of ADSCs in vitro.ADSCs in the CGF group showed higher level of ALP activity than that in the control group (P＜0.05).CGF stimulated the expression of the genes associated with osteogenesis,such as Col-I and Runx2.Conclusion:CGF can promote the proliferation and osteogenic differentiation in ADSCs in vitro.