Abstract:Malignant diseases, cerebrovascular diseases, and heart diseases have ranked the top 3 causes of death in urban China over the past decade. Due to lack of effective prevention and control method, malignant diseases are currently the major cause of death prior to the average life span in China. A focus of modern medicine is to explore the risk factors and the progression of malignant diseases. Community-based epidemiological research on malignancies lays a foundation for accurately determining the incidences, mortalities, and their changes, assessing disease burden of malignancies, exploring the etiological factors and their evolutionary rule, conducting pertinent prophylaxis, and evaluating the therapeutic efficacy. The key for etiological research of malignant diseases is to understand the dynamic interactions between environmental risk factors and the genetic susceptibility. The etiological factors and important biomarkers for early diagnosis of malignant diseases should be further verified in community-based cohort studies. Community-based population medical research possesses particular advantages in evaluating the effect of clinical interventions on survival of patients with malignancies. Enforcement of community-based population medicine is of great public health significance in reducing the incidence and improving the survival of patients with malignant diseases.

Abstract:Objective To investigate the incidence and mortality of renal cell carcinoma (RCC) patients among permanent residents in Yangpu district of Shanghai from 2002 to 2012. Methods The data of RCC patients among permanent residents of Yangpu district were collected from the database of cancer registration and management system in Shanghai. The temporal trend of RCC incidence was analyzed by using annual percent change (APC) model. Kaplan-Meier analysis with log-rank test was employed for survival analysis. The incidence and mortality were standardized with the age structure of standard population from a nationwide census taken in 2000. Results A total of 1 092 RCC cases were newly diagnosed from 2002 to 2012 in Yangpu district, and the number accounted for 2.34% of total cases diagnosed with malignant diseases. The annual crude incidence of RCC was 9.16/10⁵ and the standardized one was 5.12/10⁵ in our study, with the incidence of RCC being significantly higher in males than in females (crude incidence: 11.79/10⁵ vs 6.41/10⁵, P<0.01; standardized incidence: 6.54/10⁵ vs 3.61/10⁵, P<0.01). The incidence of RCC was significantly increased in males elder than 40 years and in females elder than 55 years. The crude incidences of RCC were annually increased in males (APC=10.45) and in females (APC=6.37) from 2002 to 2012. A total of 369 cases died of RCC during this period, with an annual crude mortality of 3.10/10⁵ and a standardized one of 1.33/10⁵. The 5-year survival rate of RCC patients was higher in males than in females (71.80% vs 69.77%, P>0.01). The 5-year survival rate of patients receiving surgical treatments was significantly higher than that of patients receiving no surgical treatments (87.75% vs 53.69%, P<0.01). Conclusion Age and gender are the main factors influencing the incidence of RCC in Yangpu district of Shanghai. The mortality of RCC is apparently lower than the incidence of RCC. Surgical treatment can effectively prolong the survival of RCC patients.

Abstract:Objective To analyze the incidence and mortality of bladder cancer patients among permanent residents in Yangpu district of Shanghai from January 2002 to December 2012. Methods The data of bladder cancer patients among the permanent residents of Yangpu district were obtained from the database of cancer registration and management system in Shanghai. The temporal trend in the incidence and mortality of bladder cancer was assessed using annual percent change (APC) model. Kaplan-Meier analysis with log-rank test was performed for survival analysis. The incidence and mortality were standardized with age composition of standard population from a nationwide census carried out in 2000. Results A total of 1 252 cases with bladder cancer were newly diagnosed between 2002 and 2012 in Yangpu district, and it accounted for 2.69% of the total patients with malignant diseases. The annual crude incidence of bladder cancer was 10.51/10⁵, with that of males being significantly higher than that of females (15.59/10⁵ vs 5.18/10⁵, P<0.01). The standardized incidence was 4.83/10⁵, with that of males being also significantly higher than that of females (7.26/10⁵ vs 2.32/10⁵, P<0.01). The crude incidence of bladder cancer showed a significant increase (APC=4.14, P<0.01) from 2002 to 2012; however, the standardized incidence showed no significant increase. The incidence increased rapidly after 50 years old. A total of 690 cases died of bladder cancer during the study period. The annual crude mortality of bladder cancer was 8.61/10⁵ in males, which was significantly higher than that in females (2.84/10⁵, P<0.01). The standardized martality was 3.64/10⁵ in males and 1.15/10⁵ in females (P<0.01). The crude mortality of bladder cancer were increased significantly and the standardized one did not alter significantly during this period. The 5-year survival rate of the newly diagnosed bladder cancer patients was 63.15%, with that of surgically treated ones being significantly higher that of non-surgically treated ones (74.31% vs 51.80%, P<0.01). Conclusion The crude incidence and mortality of bladder cancer are greatly increased during 2002-2012. Surgical treatment can significantly prolong the survival of bladder cancer patients. Early diagnosis and prompt surgical resection can improve the overall survival of bladder cancer patients.

Abstract:Objective To investigate the incidence and mortality of prostate cancer (PCa) among male permanent residents in Yangpu district of Shanghai during 2002-2012. Methods The data of PCa patients among male permanent residents of Yangpu district from January 2002 to December 2012 were obtained. Changes in the incidence and mortality of PCa was assessed using annual percent change (APC) model. Kaplan-Meier analysis with log-rank test was applied to estimate the survival. The incidence and mortality were standardized with age composition of standard male population from a nationwide census of 2000. Results A total of 1 578 cases of PCa were newly diagnosed between 2002 and 2012,accounting for 6.36% (1 578/24 800) of total male cases with malignant diseases during the period in Yangpu district, Shanghai. The annual crude incidence of PCa was 25.87/10⁵ and the standardized incidence was 10.60/10⁵. The crude incidence of PCa increased consecutively (APC=10.11, P<0.01) from 2002 to 2012; the same was true for the standardized incidence (APC=7.27, P<0.01). The incidence of PCa was increased rapidly above 60 years old. A total of 716 cases died of PCa in this period, with the annual crude mortality being 11.74/10⁵ and the standardized one being 3.92/10⁵. The mortality was increased greatly in those above 65 years old. The overall 5-year survival of the PCa patients was 57.59%. The 5-year survival rate of patients receiving operation was significantly higher than that of patients receiving no operation (67.56% vs 50.40%, P<0.01). Conclusion Age is the major reason for increased incidence of PCa in Yangpu district of Shanghai. The mortality of PCa is apparently lower than the incidence, and surgical treatment is an important factor influencing the survival of PCa patients.

Abstract:Objective To investigate the incidence and mortality of cervical cancer among female permanent residents in Yangpu district of Shanghai from Jan. 2002 to Dec. 2012. Methods The data of cervical cancer patients among female permanent residents of Yangpu district were collected from the database of the registration and management system of Shanghai. The temporal trend in the incidence and mortality of cervical cancer during this period was assessed by annual percent change (APC) model. Kaplan-Meier analysis with log-rank test was applied for survival analysis. The incidence and mortality were standardized with age composition of standard female population from a nationwide census taken in 2000. Results A total of 628 cervical cancer patients were newly diagnosed between 2002 and 2012, and it accounted for 2.53% of female malignant diseases during the period in Yangpu district. The crude incidence of cervical cancer was 10.80/10⁵ and the standardized incidence was 9.56/10⁵. The crude incidence of cervical cancer increased consecutively during the period (APC=11.10, P<0.01), whereas the standardized incidence showed no notable change (APC=1.05,P>0.05). The incidence of cervical cancer reached the top in females aged 30-49 years old. A total of 256 cases died of cervical cancer during this period; the annual crude mortality of cervical cancer in our study was 4.40/10⁵ and the standardized one was 2.02/10⁵. The average survival time of patients with cervical cancer was 8.36 years in our study. The survival time was significantly longer in the patients receiving curative surgery compared with those did not (9.22 vs 7.06 years, P<0.01).The overall 5-year survival rate of cervical cancer patients was 75.45%, with the rate of patients receiving curative surgery being significantly higher than that of those receiving no surgery (84.69% vs 60.89%, P<0.01). Conclusion In permanent female residents in Yangpu district of Shanghai, the standardized incidence and mortality rates of cervical cancer are 9.56/10⁵ and 2.02/10⁵, respectively. Surgical treatment can greatly improve the survival of cervical cancer patients.

Abstract:Objective To develop a scoring system for estimating advanced colorectal neoplasm risk in average-risk population in southern Jiangsu province, and to evaluate its screening efficiency. Methods An average-risk population of colorectal neoplasm in southern Jiangsu province, who underwent colonoscopy, was included in this study. All participants were asked to complete a questionnaire on demographic characteristics, medical history, smoking, alcohol consumption, dietary intake, and other factors that may be associated with advanced neoplasms. A multivariable logistic regression method was used to identify independent predictors of advanced neoplasms. A scoring system was developed from the logistic regression model by using a regression coefficient-based scoring method, and then was internally validated. The screening efficiency of the scoring system was assessed by its calibration, discrimination, and accuracy. Results A total of 905 average-risk participants were included in this study. The scoring system comprised 5 variables (age, sex, coronary artery disease, egg intake, and defecation frequency), with scores ranging from 0 to 10. The system had good calibration (P=0.205) and good discrimination (area under the receiver operating characteristic curve = 0.75, 95% confidence interval: 0.69-0.82). If score 2.5 was used as the screening cut-off value, the sensitivity, specificity, accuracy rate, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio were 93.8%, 47.6%, 50.1%, 9.1%, 99.3%, 1.79, and 0.13, respectively. Among the participants with low-risk (0-2) or high-risk (>2) scores, the risks of advanced neoplasms were 0.7% and 9.1% (P<0.001), respectively. If colonoscopy was used only for persons with high risk, 93.8% of persons with advanced neoplasms would be detected while the number of colonoscopies would be reduced by 45.4%. Conclusion The scoring system in this study has satisfactory screening efficiency and can be used for preliminary screening of advanced colorectal neoplasms in average-risk population in southern Jiangsu Province.

Abstract:Objective To determine the prevalence of human cytomegalovirus (HCMV) infection and plasma level of its neutralizing antibody in hypertension patients, so as to discuss the association between HCMV infection and hypertension. Methods A total of 51 anti-coagulated blood samples were collected from hypertension patients and the 50 control samples were obtained from age- and sex-matched healthy controls in the Second People’s Hospital of Anhui Province from Jan. 2013 to Apr. 2013. Neutralizing antibody level was analyzed by rapid fluorescence micro-neutralization; plasma anti-HCMV IgG and anti-HCMV IgM were determined by enzyme-linked immuno sorbent assay (ELISA), and HCMV UL93 DNA was detected by nested PCR. Results The positive rates of HCMV UL93 DNA and anti-HCMV IgG in hypertension patients were significantly higher than those in healthy controls (72.55% vs 56.00% for UL93 DNA and 70.59% vs 50.00% for IgG, P<0.05). The positive rates of anti-HCMV IgM were not significantly different between the two groups (3.92% vs 2.00%, P>0.05). The geometric mean titer of HCMV neutralizing antibody was 38.56±20.42 in hypertension patients, which was significantly lower than that in healthy controls (60.12±25.38, P=0.0326). Conclusion HCMV positive rate is higher in hypertension patients than in healthy controls; however, the specific neutralizing antibody level of HCMV is greatly lower than that in healthy controls, suggesting that the humoral immune status is declined in hypertension patients and there is a correlation between HCMV infection and hypertension.

Abstract:Objective To explore whether Antrodia cinnamomea can prolong the survival time of stroke-prone spontaneously hypertensive rats, and to investigate the underlying mechanisms from the proteomics perspective. Methods A total of 80 male stroke-prone spontaneously hypertensive rats were randomly divided into two groups (control group and Antrodia cinnamomea treated group, n=40). The animals were intragastrically given Antrodia cinnamomea (150 mg/kg). Then the death of the animals in the two groups was observed. Another 6 stroke-prone spontaneously hypertensive rats were randomly divided into drug treatment group and control group, with the drug treatment group intragastrically given Antrodia cinnamomea (150 mg/kg), and the brain proteomics were compared between the two groups 90 days later, with WKY rats used as normal controls. The differentially expressed proteins before and after drug treatment were subjected to mass spectrometry analysis, and the results of mass spectrometry analysis were further confirmed by Western blotting analysis. Results Antrodia cinnamomea significantly prolonged the survival time of stroke-prone spontaneously hypertensive rats (P<0.05), and proteomics results showed that Antrodia cinnamomea increased the expression of glutathione S-transferase (GST) and superoxide dismutase (SOD), which were also confirmed by Western blotting analysis. Further study revealed that the total anti-oxidative capability (T-AOC) of the animals was increased (T-AOC: [66.48±16.17] U/g vs [124.75±28.43] U/g, P<0.05), which was manifested by the increased activity of GST and SOD (GST: [40.33±5.24] U/mg vs [70.50±6.24] U/mg, P<0.05; SOD: [109.25±23.61] U/mg vs [192.60±23.95] U/mg, P<0.05), and decreased level of malondialdehyde ([3.96±0.45] nmol/mg vs [2.04±0.31] nmol/mg, P<0.05). Conclusion Long-term treatment with Antrodia cinnamomea can prolong the lifespan of stroke-prone spontaneously hypertensive rats, which might be related to the increased activity of anti-oxidative enzymes and the decreased oxidative damage.

Abstract:Objective To investigate the effects of S100A6 gene silencing on the proliferation and migration of Eca109 human esophagus cancer cells. Methods The shRNA expression vectors were constructed using the shRNA sequences designed based on human S100A6’s coding sequence, and were transfected into Eca109 cells via cationic liposome. The changes of S100A6 mRNA and protein in Eca109 cells transfected with the recombinant vectors were detected using real-time PCR and Western blotting analysis 48 hours after transfection, respectively; the proliferative curves of transfected cells were plotted using MTT assay; furthermore, the change in cellular migration ability was determined using wound healing assay. Results The eukaryotic expression vector of shRNA targeting S100A6 was successfully constructed. Real-time PCR and Western blotting analysis results showed that S100A6 was effectively silenced by liposome-mediated transfection of the recombinant shRNA vectors in Eca109 cells. Compared with the untransfected cells, S100A6 mRNA and protein in transfected Eca109 cells were significantly decreased (P<0.05, P<0.01). Meanwhile, the proliferative activity of Eca109 cells was significantly inhibited by S100A6 silencing (P<0.01). It was found that the cellular migration was also suppressed by S100A6 gene interference. Conclusion S100A6 gene can be effectively silenced by shRNA expression vectors, and the silence may lead to inhibition of the proliferation and migration of Eca109 cells.

Abstract:Objective To evaluate the effect of the galacto-oligosaccharides (GOS) on the intestinal mucosal tight junction and barrier function of rats with induced cold restraint stress. Methods Forty Wistar rats were randomly divided into four groups: two cold resistant stress (CRS) groups fed with either a standard diet (CRS group) or 4 g/kg GOS per day (CRS+GOS group) and two sham CRS groups fed with a standard diet (control group) or 4 g/kg GOS per day (GOS group). All the four groups were fed for a total of 10 days, and the CRS and CRS+GOS groups were subjected to cold restraint stress on the 10^th day. Then the blood samples of all the four groups were collected via cardio puncture for determination of serum diamine oxidas (DAO) activity; finally all the rats were sacrificed and the intestinal tissues were collected for pathological and electronic microscopic examination, with the occludin mRNA level measured by real-time PCR and the occludin protein expression by immune-histological method. Results Compared with the control, the serum DAO levels of CRS group and the CRS+GOS group were significantly increased(P<0.01); and that in the CRS group was significantly higher than that in the CRS+GOS group (P<0.01). The levels of occludin mRNA and protein expression were significantly decreased in the two CRS groups (P<0.01), and that in the CRS group was significantly lower than that in the CRS+GOS group (P<0.01); the serum DAO levels, the expressions of occludin mRNA and protein were not significantly different between the control and the GOS groups. Conclusion Feeding with GOS can decrease plasma level of DAO in rats, increase expression of intestinal occludin mRNA and protein in rat ileum, indicating that GOS can improve the mucosal tight junction and intestinal barrier function.

Abstract:Objective To investigate the fragmentation pathways of sildenafil and its active metabolite N-desmethylsildenafil, so as to understand the intestinal absorption and metabolic characteristics of sildenafil. Methods Using electrospray ion technology (positive ion mode), we analyzed the fragmentation pathways of sildenafil and its active metabolite N-desmethylsildenafil. Based on the rule of cracking, a LC-ESI(+)MS/MS method was developed to determine the intestinal absorption of sildenafil and metabolism of active metabolites N-desmethylsildenafil in the mesenterium. Totally 25 healthy adult male SD rats were randomly divided into five groups and each group contained 5 rats. The hepatic portal venous blood samples were obtained from rats after intestinal administration of 10 mg/kg of sildenafil citrate. The absorption and metabolism were examined in the in situ intestinal administration model at premedication, 0.25 h, 0.5 h, 1 h and 4 h (five groups) after medication. Results Mass spectrometric pyrolysis of sildenafil and its active metabolite N-demethylsildenafil showed ion peaks m/z 311 and 283, indicating that the C-S bond was unstable and was liable to lose C₅H₁₂O₂N₂S, and the C-O bond was unstable and was liable to lose C₂H₄, finally forming stable fragment ions. When m/z 475→m/z 283 (sildenafil) and m/z 461→m/z 283 (N-demethylsildenafil) were used as ion reaction channel, sildenafil were absorbed not only in the form of sildenafil in the intestine, but also in the form of its active metabolite N-demethylsildenafil metabolisming as 1/5 of the prototype drugs were finally changed into N-demethylsildenafil. Conclusion Both sildenafil and N-demethylsildenafil can be cracked into stable m/z 283 secondary ions. Intestinal absorption is accompanied by its nitrogen demethylation reaction, which leads to the fact that about 1/5 of prototype drug metabolized into active metabolite N-demethylsildenafil.

Abstract:Objective To induce synchronous differentiation of rabbit bone marrow-derived mononuclear cells into endothelial progenitor cells (EPCs) and smooth muscle progenitor cells (SPCs), and to study their biological properties and the possibility of them as seed cells for tissue-engineered venous valves. Methods Gradient density centrifugation was used to obtain bone marrow blood mononuclear cells, which were separately cultured with EGM-2 complete medium containing 5% FBS for differentiation of EPCs and with EBM-2 medium without vascular endothelial growth factor (VEGF) containing 5% FBS and 20 ng/mL platelet-derived growth factor-BB (PDGF-BB) for differentiation of SPCs. The differentiation of EPCs and SPCs was identified by various methods. Results EPCs were cultured for 10 days and the cells fused into monolayer, showing a “stepping stone” appearance and expressing VEGF receptor-2 (VEGFR-2), von Willebrand factor (vWF) and CD133, but not α-smooth muscle actin (α-SMA); Weibel-Palade bodies were seen within the EPCs cytoplasm under the transmission electron microscope. Biological function tests showed visible EPCs growing on the matrigel in a blood vessel-like form. SPCs were cultured for 14 days and showed the specific features of the vascular smooth muscle growth, namely, the “peak-valley” growth way. SPCs expressed CD34 and α-SMA but not vWF and VEGFR-2. Myofilaments, paralleling with the cell longitudinal axis, were seen under the transmission electron microscope. SPCs could not form vessel-like structures on the matrigel. Conclusion Mononuclear cells can be obtained through gradient density centrifugation of the bone marrow blood, which can be synchronously induced into EPCs and SPCs, providing economical and easy seed cells for tissue-engineered venous valves.

Abstract:Objective To investigate the correlation of suicidal ideation with social support and quality of life (QOL)among college students. Methods A total of 9 808 college students in Chongqing were surveyed by using stratified cluster random sampling method to investigate their suicidal ideation status; the association of suicidal ideation with social support and QOL was analyzed by multipleregression correlation analysis. Results The reporting rate of suicidal ideation was 13.04% in our survey,especially high in females (16.39%), art students (15.09%), students with religious belief (15.05%), and students with history of suicide attempt (68.64%). The students with suicidal ideation had lower scores in social support,objective support,subjective support, utilization of support, QOL, physical condition, psychological health,economic status, working condition, family relationship and interpersonal relationship with others than students without suicidal ideation (P<0.01). It was also found that the suicidal ideation was negatively correlated with subjective support (r=-0.100, P<0.01), utilization of support (r=-0.094, P<0.01), physical condition (r=-0.127, P<0.01), psychological health (r=-0.067, P<0.01), and economic status (r=-0.081, P<0.01). Conclusion College students in Chongqing have a high prevalence of suicidal ideation. Society, family and school should attach more importance to the mental health status of college students and improvement of social support system for college students; comprehensive measures should be taken to prevent suicidal ideation and behavior.

Abstract:Objective To evaluate the efficacy of glucocorticoids in treatment of patients with severe acute pancreatitis (SAP). Methods According to the inclusion criteria, the experimental design, characteristics of the included objects, and the corresponding outcomes were extracted from relevant literatures. Meta-analysis was done to evaluate the results by RevMan 5.1 software. Results A total of eight articles with 184 cases in this treatment group and 177 participants in the control group were included in this meta-analysis. Data from six studies showed that the mortality rate of glucocorticoid group (14/145,9.655%) was significantly lower than that of the control group (28/139,20.14%), with the difference being statistically significant (OR=0.42,95% CI [0.21-0.83], P=0.01). Data from three studies showed that the frequency of acute respiratory distress syndrome (ARDS) of the glucocorticoid group (20/95,21.05%) was significantly lower than that of the control group (37/89,41.57%, OR=0.33,95% CI [0.16-0.65],P=0.001). Data from three studies showed that the operation transferring rate in the glucocorticoid group (12/87, 13.79%) was significantly lower than that of control group (26/81,32.09%, OR=0.29, 95% CI [0.13-0.67], P=0.004). Compared with the control group, the glucocorticoid group also had significantly reduced the mean hospital stay in four experiments (standardized mean difference -1.52, 95% CI [-2.73--0.32],P=0.01). Conclusion Glucocorticoid has advantages over conventional treatments in reducing the incidence of ARDS and mortality and shortening hospital stay in SAP patients, which provides a new and prospective therapeutic option for SAP.

Abstract:Objective To evaluate the effect of chemotherapy on detection of colorectal cancer autoantibodies by phage display peptide method. Methods The five-phage peptide clones (No.95, No.149, No.174, No.396, and No.1009) with high discriminatory ability of colorectal cancer patients were selected as the study subjects. We compared the reactivity of autoantibodies against each of the five-phage peptide clones among 20 colorectal cancer patients receiving chemotherapy, 40 colorectal cancer patients receiving no chemotherapy, and 40 healthy controls by enzyme linked immunosorbent assay (ELISA). Results The seroreactivity of No.95 clone was significantly lower in the patients with chemotherapy than those without chemotherapy (P<0.05), and was similar between normal control group and patients with chemotherapy (P=0.074). For the seroreactivity of the other four clones (No.149, No.174, No.396, No.1009), there were significant differences between patients with chemotherapy and normal controls (all P<0.01). Conclusion The detection ability of No.95 clones for colorectal cancer autoantibodies is greatly influenced by chemotherapy, while chemotherapy shows no notable influence on the reactivity of other four-phage peptide clones.

Abstract:Down’s syndrome is an autosomal disease and its prenatal screening is particularly important. In recent years, clinical application of serological screening markers has been widely used, which improved the detection rate of Down’s syndrome, but still remained at about 60%. The new generation of sequencing technology for direct analysis of nucleic acids, which can be used for fetal chromosomal aneuploidy detection, can detect Down’s syndrome more accurately. In this paper, we compared the prenatal screening methods for Down’s syndrome in recent decades, and reviewed the new screening markers based on proteomics and bioinformatics technology for clinical application.

Abstract:K-complex (KC) is a major component of electroencephalography (EEG) in human brain during non-rapid eye movement sleep, and there has long been a debate on its function. Some researchers consider that KC represents a sign of arousal response of brain to internal or external stimuli, and others suggest that it is a protective response to sustain sleep. In this article we put forward a functional model of KC based on the above two views and related literatures. According to our model, KC arises from three kinds of brain rhythms (sleep cyclic, central oscillation, and cortical oscillation). Based on the results of stimulus significance assessed by sleep-information assessment center, the generation of KC is related to three different responses-total arousal, sign of being awakened, and no sign of being awakened. In conclusion, KC is a reflection of cerebral state responsive to stimuli.

Abstract:Objective To compare the efficacies of three methods (Qiagen kit, Tiangen kit and modified manual extraction) for extracting human genomic DNA from fresh and frozen blood samples. Methods Twenty samples of intravenous whole blood (5 mL/sample) were collected from pancreatic cancer patients and centrifuged. Each sample was then divided into fresh and frozen samples. Human genomic DNA was extracted from the fresh blood samples within 12 h by the three methods: Qiagen kit, Tiangen kit and modified manual extraction. The frozen sample was thawed to room temperature after preserved at -40℃ for 72 h and the DNA was extracted by the same three methods. The integrity of the DNA products was examined by agarose gel electrophoresis; the purity and concentration were examined by nucleic acid protein detector; and the cost and time spent of the 3 methods were evaluated. Results The DNA integrity by the modified manual method was better than those of the other two methods using kits. As for purity, some frozen samples presented ghost peaks of contamination. As for the yield of the DNA products, the Qiagen kit was superior to Tiangen kit for both fresh and frozen samples. The yield of modified manual method was similar to that of Tiangen kit for frozen samples, and was the lowest one for fresh samples. Methods using kit took less time but cost more. Conclusion The modified manual extraction can obtain longer and more intact genomic DNA compared with the other two methods. Although the modified manual extraction is inferior in yield and time saving, it costs less and is more suitable for genomic DNA extraction from large frozen samples.

Abstract:Objective To study the interaction between biomacromolecules (protein and DNA) and different Rhizoma Coptidis alkaloids (palmatine, coptisine, epiberberine, jatrorrhizine and berberine) and the difference among 5 Rhizoma Coptidis alkaloids. Methods Bovine serum albumin (0.5 mL of 1.10×10^-5 mol/L) was added to 1.5 mL of each alkaloid of different concentrations (concentrations of palmatine, coptisine, epiberberine, jatrorrhizine and berberine being 0, 1.95×10^-6, 3.9×10^-6, 7.8×10^-6, 1.56×10^-5 and 3.12×10^-5 mol/L, respectively). The reaction system was shaken and incubated at 37℃ for 1 h, and then the emission spectra of mixed solution were recorded within 300 to 500 nm at 280 nm excitation. Plasmid DNA (0.5 mL of 0.1%) was added to 1.5 mL of each alkaloid with different concentrations (concentrations of palmatine, coptisine, epiberberine, jatrorrhizine and berberine being 0, 1.95×10^-6, 3.9×10^-6, 7.8×10^-6 and 1.56×10^-5 mol/L, respectively). The reaction mixture was well shaken, and then the fluorescent emission spectrum was recorded from 480 to 650 nm at 368 nm excitation by an F-4500 fluorescence spectrophotometer. Results All the Rhizoma Coptidis alkaloids in test exhibited similar quenching effect on fluorescence of bovine serum albumin, and they all showed fluorescence enhancing effect on plasmid DNA, and the enhancing effects varied greatly among the five alkaloids, with epiberberine showing the strongest activity, followed by coptisine, palmatine, berberine, and jatrorrhizine in order. The strongest interaction between DNA and five alkaloids were found at the concentration of 7.8×10^-6mol/L. Conclusion Strong interaction can be found between Rhizoma Coptidis alkaloids and biomacromolecules including protein and DNA. The interaction of Rhizoma Coptidis alkaloids with plasmid DNA varies greatly among different alkaloids and is similar with bovine serum albumin. 9,10-methylene-dioxy groups and 2,3-dimethoxy groups might have promoted the interaction between alkaloids and DNA.

Abstract:Objective To develop a high-performance liquid chromatography (HPLC)-diode array detector (DAD) method for determination of 20 colorants in hair dyes, so as to improve the method for determining oxidative hair dyes in Hygienic Standard for Cosmetics. Methods After ultrasonic extraction with ethanol-water, samples were analyzed by HPLC. The separation was achieved on an Agilent Zorbax Bonus-RP column (4.6 mm×250 mm,5 μm) using a gradient mobile phase system of A: acetonitrile and B: 4 mmol/L PBS (pH=6.0) containing 0.1% 1-octanesulfonic acid sodium salt solution at a flow rate of 1 mL/min. The gradient elution program was as follows: 0-24 min, 10% A; 24-26 min,10%-40% A; 26-45 min,40% A; 45-47 min,40%-80% A; 47-52 min,keep 80% A unchanged. The detection wavelength was set at 280 nm,the temperature of column was 15℃, and the injection volume was 5 μL. Results All the components were separated at baseline within 50 min. Within the concentration range of 10.0-500.0 mg/L, the peak areas and concentrations of colorants had good linear relationship, with the linear correlation coefficients being higher than 0.999. The intra-day and inter-day precisions were within 4% and the average recoveries ranged from 81.90% to 119.88%. Conclusion The present method is simple, rapid and accurate and is suitable for determination of colorants in oxidative hair dyes.

Abstract: