Objective. We aimed to investigate the relationship between FGF21 and obesity-related acanthosis nigricans (AN). Methods. 40 obese patients without AN (OB group), 40 obese patients with AN (AN group), and 40 healthy volunteers  were included in this study. Weight, BMI, lipid profile, FFA, UA, and CRP were measured in all participants. Oral glucose tolerance tests (OGTT) were performed and serum glucose and plasma insulin were measured. Serum FGF21 was measured by ELISA. Results. Compared with OB group, AN group had higher levels of fasting insulin and homeostasis model of assessment for insulin resistance (HOMA-IR) (P < 0.05), but lower serum levels of blood glucose. The difference of FGF21 among three groups was significant and AN group showed the highest serum level of FGF21 (P < 0.05). Serum FGF21 was most positively correlated with fasting insulin and HOMA-IR. Multiple logistic analysis showed that FGF21 was the independent risk factor for AN . Conclusion. AN patients had more serious hyperinsulinemia but better serum levels of blood glucose than OB. Increased FGF21 is associated with AN in obese patients and may be considered as compensatory response to the decreased insulin sensitivity. Acanthosis nigricans (AN) is characterized by hyperpigmented and papillose thickening of the epidermis, which is mainly present on the skin folds, such as the axillae, posterior neck, umbilicus, and occasionally mucosal surfaces , 2. ClinFibroblast growth factor 21 (FGF21) is a typical member of the fibroblast growth factor family. FGF21 is mainly produced in peripheral tissue such as liver, white/brown adipose, pancreas, and skeletal muscle . It servα induces FGF21 expression, which is required for the activation of hepatic lipid oxidation [Previous research showed that FGF21 plays a key role in the adaptive response to fasting. During fasting, the activated PPARxidation , 5. In dxidation –8. In roxidation , 10. FGFIn this study, we detected serum FGF21 levels in obese patients and patients with obesity-related AN. Our data demonstrated that compared with obese patients, FGF21 levels were positively correlated with obesity-related AN and markers of insulin resistance .n = 40, BMI > 28 kg/m2), obesity with AN , and normal controls . Our study was approved by the hospital ethics committee and the clinical trials registration number is ChiCTR-OCS-12002381, and all the participants were asked to sign an informed consent prior to participation in the study. Participants with severe systemic disease were not included in this study.40 obesity-related AN patients were admitted to our department from December 1, 2013, to August 31, 2014. 40 BMI-matched simple obese patients and 40 healthy volunteers (age from 20 to 31 years) were also included in the study. All the participants were aged >18 years and were divided into three groups: simple obesity ; 3—moderate: extending to the lateral margins of the neck ; it should not be visible when the participant is viewed from the front; and 4—severe: extending anteriorly (>6 inches), visible when the participant is viewed from the front. Axilla severity was as follows: 0—absent: not detectable on close inspection; 1—present: clearly present on close visual inspection, not visible to the casual observer, extent not measurable; 2—mild: localized to the central portion of the axilla; it may have gone unnoticed by the participant; 3—moderate: involving entire axillary fossa, but not visible when the arm is against the participant's side; and 4—severe: visible from front or back in the unclothed participant when the arm is against the participant's side. In this study, each subject enrolled with AN had a score greater than 2. Each subject had completed secondary or higher education.The following scale for AN was used . Neck seBody parameters and biochemical parameters, weight subjects (kg), height (cm), body fat, and body mass index (BMI), were measured by simple anthropometric measuring instrument  by professional physician and also percentage of body fat, visceral fat fraction, basal metabolic rate, and body age were calculated at the same time. Morning fasting venous blood was collected. Total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL), free fatty acids (FFA), uric acid (UA), C reactive protein (CRP), and thyroid stimulating hormone (TSH) in serum were detected.The homeostasis model of assessment for insulin resistance (HOMA-IR) was calculated on the basis of fasting values of plasma glucose and insulin according to the HOMA model formula: HOMA-IR = fasting insulin × fasting glucose, divided by 22.5 . Serum Fx  ±  SD) and the count data as the number of columns (n). Student's t-test and chi-square tests were used for statistical comparison. Pearson or Spearman method was used for correlation analysis to assess the correlation between different variables. Multivariable logistic-regression analyses were performed to determine the risk factors for AN in obese patients. P values were considered to be significant below 0.05 (P < 0.05).All statistical tests were performed using SPSS18.0 software, and quantitative data were expressed as mean ± standard deviation (P < 0.05), and AN group had higher level of FFA than OB and CON groups (P < 0.05). Obesity and AN were associated with lipids dysfunction and inflammation.P < 0.05) than the CON group. Additionally, the OB group had higher blood glucose levels than AN group at each time point although the difference is not significant. Compared with the CON group, the OB group showed higher levels of insulin at four time points  (P < 0.05), and the AN group had higher levels at each point (P < 0.05). Compared with the OB group, the AN group had higher levels of insulin at three time points  (P < 0.05) . The results in  < 0.05) . Compare < 0.05) . HOMA-IR < 0.05) . These rP = 0.042), while increased fasting insulin was associated with AN but failed to reach statistical significance .We speculated that AN could affect serum FGF21 levels. The results shown in Obesity is a public health problem that has raised concern worldwide . Acantho2, P < 0.001), while, in our study, BMI of AN and OB patients was similar . In addition, the higher level of FGF21 is compensatory for the insulin resistance and is response to downregulation of blood glucose.The pathogenesis of AN was closely associated with insulin resistance –16, whilβ cells function and promote glucose uptake, which could explain the reduced blood glucose levels in AN patients. In addition, studies have shown that FGF21 can promote hepatic gluconeogenesis and the consumption of fat, increase lipid metabolism and fat utilization, and also promote the conversion of FFA to ketone body [Serum FGF21 levels are elevated in patients with obesity, type 2 diabetes, and nonalcoholic fatty liver disease (NAFLD)  and coulone body , 25. Theone body . MoreoveThe relationship between FGF21 and obesity remains controversial –32. AfteIn summary, the main finding of our study was that serum FGF21 was increased in response to insulin resistance in AN. FGF21 may play an important role in the pathogenesis of AN. However, our study also has limitations, such as the relatively small number of patients and the mechanism of how FGF21 participates in the AN is yet to be fully elucidated. Larger studies with more patients are required to confirm the association between FGF21 and insulin resistance in AN. More mechanistic studies are needed to define the signaling mechanisms linking FGF21 with insulin resistance.


Subgroup analysis revealed that time lag, study location, number of patients, proportion of females, study design, number of adjusted variables might be potential source of heterogeneity. Sensitivity analysis showed robustness of the results. No publication bias was observed in studies on AHD or LP. The current meta-analysis indicated that males are at higher risk of AHD or LP compared with females. More attention should be paid to males to make sure early testing, diagnosis, and treatment, and ultimately improve individual and population health.Effectiveness of highly active antiretroviral therapy is limited for a large proportion of individuals living with HIV presenting for medical care at an advanced stage. Controversial results of gender differences in risk of late HIV diagnosis were reported among existing literatures. Therefore, we conducted this meta-analysis to synthesize a summary of gender differences in risk of advanced HIV disease (AHD) and late presentation (LP) according to European consensus definitions. Totally, 32 studies were included based on predetermined selection criteria. The pooled adjusted  Highly active antiretroviral therapy (HAART) has dramatically reduced HIV-related morbidity and mortality since it has been available in the mid 1990s. Many HIV infected individuals are living with HIV/AIDS as a chronic manageable disease rather than an inevitably progressive and fatal illness13Late presenters are associated with higher rates of opportunistic infections578610111415171820However, different criteria have been used to define late presentation among HIV infected individuals, which generally include CD4 cell count and/or AIDS-defining diseases12Previous studies showed that males were more likely to present late in HIV diagnosis252795% confidence interval (CI), 33 articles with inconsistent definition of outcomes, 3 articles only recruited patients infected via heterosexual contact, 2 articles contained unspecified the exact duration between initial HIV diagnosis and AIDS diagnosis, were excluded. After this selection, only one study used Poisson regression adjusted by age as a quadratic to calculate adjusted relative risk (aRR), while the others used logistic regression to calculate adjusted odds ratios (aOR). This study was finally excluded owing to small number and inconsistency in method, leaving 32 studies for final inclusion in the meta-analysis.The search strategy retrieved 1095 unique citations. Of these, 935 citations were excluded after the first screening based on abstracts or titles, leaving 160 articles for full-text review . On thisCharacteristics of the 43 arms in 32 selected studies are shown in aORs were reported in all studies. All risk measures were adjusted for confounding factors with number of adjusted variables ranging from 2 to 16.The selected studies were published between 2002 and 2014. The number of participants per study/arm on advanced HIV diseases ranged from 113 to 28382, for a total of 1.55 million participants across 29 arms (63734 incident cases of AHD). The number of participants per study/arm on late presentation ranged from 352 to 5545, for a total of 48923 participants across 12 arms (29754 incident cases of LP).Nine studies were conducted in the United States, 8 in Italy, 3 in Spain, 2 in France, 2 in Mexico, and the other 10 studies were conducted in the following countries or region: Hong Kong, Thailand, India, Vietnam, Germany, Nigeria, Republic of Korea, Belgium. The percentage of females ranged from 9.00% to 70.88%. The et al., 2010a and Yang et al., 2010b were excluded to calculate the pooled aOR due to shorter time lag), all but two found higher risk of presenting with AHD for males compared with females, although not all were statistically significant. Males had an increased risk of presenting with advanced HIV disease compared with females, with a pooled aOR of 1.73    to 1.76  for studies on AHD  to 1.43  for studies on LP (Among the 29 arms in 26 selected studies on AHD (Yang 59–1.89) . Among t18–1.62) . A sensis on AHD , and 1.3es on LP .aOR of 1.85  with 0 month, aOR of 1.53  with 1 month, aOR of 1.72  with 3 month, aOR of 1.67  with 6 month, and aOR of 1.70  with 12 month, respectively. The pooled risk of LP in males compared with females of different time lags were aOR of 1.36  with 0 month, aOR of 1.41  with 3 month, and aOR of 1.54  with 12 month, respectively. The selected study characteristics did not seem to significantly influence the results (P > 0.05), and only partly explained the source of heterogeneity.To explore the between-study heterogeneity, we performed subgroup analyses and meta-regression across a number of key study characteristics . The finz = −0.49, P = 0.626; Egger’s test: bias = 0.87, t = 1.47, P = 0.153) or LP . Visual inspection of the Begg funnel plot revealed mostly symmetrically distribution confirming the absence of publication bias  and misclassification attributable to other health care processes with a longer window I2 = 68.70%, P < 0.001) for AHD and no heterogeneity  for LP, possibly because more complex confounding factors were introduced into larger number of patients. Difference in data source and method of collecting data among cross-sectional study, retrospective study, and prospective study, might contribute to heterogeneity as well. More studies included higher proportion of males, and the different sex ratio of patients might cause heterogeneity of the pooled risk of advanced HIV disease or late presentation for males compared with females. Taken together, the result of subgroup analysis revealed that time lag, study location, number of patients, proportion of females, study design, number of adjusted variables might constitute source of heterogeneity, but only partly explained the heterogeneity.However, the proposed definitions did not specify the time lag between initial diagnosis of HIV infection and AIDS. Then Michael Kozak aOR may be a source of publication bias, despite the fact that no evidence of publication bias was exhibited by Begg’s or Egger’s tests.To the best of our knowledge, this is the first meta-analysis to quantify the risk of AHD and LP for males compared with females. Advantages of the current study are applying European consensus definitions of AHD and LP and stratifying the results by varied time lags between initial diagnosis of HIV infection and AIDS. However, several limitations should be noted. To begin with, significant heterogeneity was observed in overall and subgroup analyses. Otherwise, the observation of heterogeneity should not reduce the confidence in the finding but just add some uncertainty about the magnitude of that54In conclusion, the current meta-analysis indicates that males are at higher risk of AHD or LP compared with females. Considering the consistent findings of a number of studies, and the reliability and robustness of our meta-analysis, we strongly recommend that more attention should be paid to males, more effort should be made to encourage individuals with high-risk behavior to participate counseling and testing, in order to make sure early testing, early diagnosis, and early treatment, ultimately to improve individual and population health.This meta-analysis was conducted in accordance with the Meta-Analysis of Observational Studies in Epidemiology guidelines. Pubmed, Embase, and Web of Science databases were searched for English-language publications dated until April 30th, 2015. Keywords used in the databases search included: Search Search (((((((gender) OR sex) OR males) OR females) OR men) OR women)) AND ((((((((((“HIV testing”) AND late)) OR ((“delayed diagnosis”) AND HIV)) OR “late HIV diagnosis”) OR ((“late presenters”) AND HIV)) OR “advanced HIV disease”) OR ((“late diagnosis”) AND HIV)) OR ((“late presentation”) AND HIV)) OR ((“late testers”) AND HIV)). Chinese Scientific Journals Fulltext Database (CQVIP), China National Knowledge Infrastructure (CNKI) and Wanfang Data were searched for Chinese-language publications dated until April 30th, 2015. Keywords used in the databases search included: (“Ai Zi Bing (AIDS)” OR “HIV”) and (“Jian Ce (Testing)” OR “Zhen Duan (Diagnosis)”). References lists of the retrieved articles and previous systematic reviews were also reviewed by researchers.aOR, or aRR or adjusted Hazard Ratio (aHR) with the corresponding 95% CI of gender difference in “LP” or “AHD”. Studies were excluded if they did not specified the exact duration between initial HIV diagnosis and AIDS diagnosis. If multiple publications reported results based on the same research, the more recent or complete article was included.Two authors examined the titles and abstracts of all studies containing keywords independently for eligibility. Studies were included based on the following criteria: (1) reported original data; (2) described “LP” as presence of an AIDS condition or CD4 cell count <350 cells/μL at presentation for care, or “AHD” to as the presence of either an AIDS condition or a CD4 cell count <200 cells/μL at presentation12aOR or aRR or aHR with the corresponding 95% CI, and adjusted variables. Number of patients and cases was the number of subjects included in the model of multivariate analysis.Data extraction was carried on by two authors independently using a pre-designed data extraction form. The following information was extracted from the included publications: last name of the first author, publication year, study period, study population, study location, outcome definition, time lag , data source, study design, number of patients, number of cases (patients with late presentation or advanced HIV disease), 95% CI of gender difference in risk of AHD or LP were used to calculate the pooled estimates based on weighted pooled measures. If multiple arms using varies time lags in one study, arm with the longest time lag was included to calculate the overall pooled estimates. Forest plots were used to visually assess the individual and pooled estimates with the corresponding 95% CI, and heterogeneity among studies was assessed using the Cochrane Q statistic significant when P < 0.10 and the I2 statisticP < 0.10 and I2 < 50%58P < 0.05 was considered significant, except where otherwise specified.The adjusted estimates with the corresponding How to cite this article: Jiang, H. et al. Gender difference in advanced HIV disease and late presentation according to European consensus definitions. Sci. Rep.5, 14543; doi: 10.1038/srep14543 (2015).


Women of refugee background from Africa are reported to have a greater risk of adverse pregnancy outcomes compared to women born in resettlement countries. However, there is currently little insight into whether adverse pregnancy outcomes are more common among migrant women of refugee background, compared to women who have migrated for non-humanitarian reasons. To inform whether women of refugee background require additional services in pregnancy compared to non-refugee migrant women from similar world regions we aimed to describe and compare maternal health, pregnancy care attendance and pregnancy outcomes among migrant women from Africa with or without a refugee background.Retrospective, observational study of singleton births at a single, metropolitan, maternity service in Australia 2002–2011, to women born in humanitarian source countries (HSC) and non-HSC from North Africa (n = 1361), Middle and East Africa (n = 706) and West Africa (n = 106).Compared to non-HSC groups, age < 20 years (0–1.4% vs 2.3-13.3%), living in relatively socio-economically disadvantaged geographic areas (26.2-37.3% vs 52.9-77.8%) and interpreter need (0–23.9% vs 9.7-51.5%) were generally more common in the HSC groups. Compared to non-HSC groups, female genital mutilation (0.3-3.3% vs 5.1-13.8%), vitamin D insufficiency (8.7-21.5% vs 23.3-32.0%), syphilis (0–0.3% vs 1.2-7.5%) and hepatitis B (0–1.1% vs 1.2-18%) were also generally more common among the HSC groups. Unplanned birth before arrival at the hospital (3.6%) was particularly high in the North African HSC group. HSC-birth was associated with gestational diabetes mellitus  among women from Middle and East Africa, after adjusting for maternal age, parity, body mass index and relative socio-economic disadvantage of area of residence. The West African HSC group had the highest stillbirth incidence (4.4%).Migrant women of refugee background from different African regions appear to be at greater risk of specific adverse pregnancy outcomes compared to migrant women without a refugee background. Awareness of differing risks and health needs would assist provision of appropriate pregnancy care to improve the health of African women and their babies. Complete data for age, parity, BMI and IRSD were available for 961 women from North Africa, 491 from Middle and East Africa and 82 from West Africa. Women who were missing data regarding BMI and/or IRSD were excluded from any multivariable analysis that included these variables. Categories with low numbers were collapsed as appropriate. Fewer confounders were included if number of cases was low to avoid over-fitting.General health and demographic characteristics of the study populations are summarised in Table North African non-HSC included Algeria, Egypt, Libya, Morocco and Tunisia (n = 214); the only HSC was Sudan (n = 1147).Compared to the North African non-HSC group, mean age was lower in the North African HSC group  and a larger proportion of the HSC group were aged <20 years ; HSC included Democratic Republic of the Congo, Burundi, Eritrea, Rwanda and Tanzania (n = 87).Compared to the Middle and East African non-HSC group, mean age was greater in the Middle and East African HSC group  but there was no significant difference in median BMI (25.0 (7.0) vs 25.0 (7.0), p = 0.54). Compared to the non-HSC group, larger proportions of the HSC group were multiparous, lived in a relatively socio-economically disadvantaged area, required an interpreter, or had FGM ; HSC included Guinea, Liberia, Mauritania and Sierra Leone (n = 45).Compared to the non-HSC group, women born in HSC in West Africa had a lower mean age , a smaller proportion were 35 years or older and a larger proportion were aged less than 20 years  in women from HSC. Late first pregnancy care visits at the hospital were common in all groups. We present novel data suggesting specific HSC groups may be at greater risk of particular adverse pregnancy outcomes compared to non-HSC groups from similar world regions, such as BBA in the North African HSC group and GDM in the Middle and East African HSC group. Stillbirth was most common in the West African HSC group but was also more common than the national figures in most groups.Compared to the non-HSC groups, poorer general health , social disadvantage  and FGM were generally more common in the HSC groups. Many of these are associated with adverse pregnancy outcomes ,21-24. TEarly pregnancy care provides an opportunity to improve maternal health, connect women with social support and monitor maternal risk factors for adverse pregnancy outcomes. Pregnancy care attendance and timing of first pregnancy care visit at the hospital did not differ between non-HSC and HSC groups within any region. However, late first pregnancy care visits were more common than nationally 22-35%) in each group  and more-35% in eWhile pregnancy outcomes were not universally poorer among women of refugee background, each HSC group appeared to have high incidences of specific adverse outcomes. Our findings are consistent with previous research comparing African-born women to resettlement country-born women and advance knowledge by indicating additional risk associated with refugee background, above that associated with migrant background. GDM was independently associated with HSC-birth among women from Middle and East African countries and was more common than previous reports for African populations in Australia (4-10%) ,33. In tPerforming an episiotomy in women who have experienced FGM is sometimes used as a strategy to reduce the risk of severe perineal trauma ,36,37. IWomen from Sudan were less likely to have non-cephalic presentation, caesarean section or assisted vaginal birth than the North African non-HSC group. Further investigation revealed that a larger proportion of women from Sudan had emergency caesarean sections and a smaller proportion had repeat caesarean sections. This is consistent with research in which a mostly Sudanese group of women in Australia was less likely to have elective caesarean sections than the hospital population . CaesareAfrican migrant women have previously been reported to be less likely to use analgesia during labour than resettlement country-born women ,29,40. OBBA was most common among women from Sudan with an incidence considerably higher than nationally (0.5%) . BBA hasThere was evidence of poor neonatal outcomes in both HSC and non-HSC groups. Stillbirth incidence did not significantly differ within any region but was higher than the national rate (0.7%)  in five This study has limitations including that potential cultural and language barriers may contribute to over- or under-representation among wellbeing factors reliant on self-report e.g. spousal relationships. Some variables were only routinely collected from partway through the study time period (e.g. BMI); consequently sample size was reduced for these variables. Country of birth was used as a proxy for refugee background, however, in accordance with our previously published work , humanitThis study also has a number of strengths. It employed a rigorous, objective method to select current HSC and made the unique comparison to probable economic migrants from the same world regions. It described pregnancy outcomes for several African regions at a single maternity service, including West Africa which has not previously been described in detail. The large sample size enabled multivariable analysis to control for some potential confounders.In this unique study from one of the largest maternity services in Australia comparing women from three African regions, we have advanced knowledge on maternal health, pregnancy care attendance and pregnancy outcomes. It highlights the need for engagement with communities to improve early pregnancy care attendance as, within each African region, women born in HSC may be at greater risk of specific adverse pregnancy outcomes compared to women born in non-HSC. Early pregnancy care may also be especially important for women from West African non-HSCs who may have high rates of caesarean section and adverse neonatal outcome, although confirmatory research is needed. Awareness of these differing health needs would assist provision of appropriate care to improve the health of African women and their babies.


Animals were given a daily gavage treatment of OJS in daily dosages of 0, 500, 1000 or 2000 mg/kg for 4 weeks. Cytotoxicity assay was conducted at various concentrations in 23 different cell lines including neuroblastoma, glioblastoma, hepatocarcinoma, melanoma, leukemia, colon cancer, breast cancer, keratinocytes, phechromocytoma, prostate cancer, bronchial epithelial cells, and gastric adenocarcinoma.OJS did not induce significant changes in mortality, food consumption, organ weights, hematology, serum biochemistry, and urinalysis, except for decrease in number of white blood cells over 1000 mg/kg/day female group. Thus, the no observed adverse effect level (NOAEL) is more than 2000 mg/kg/day for male and 500 mg/kg/day for female rats. In addition, OJS had no cytotoxicity against all tested cells.Collectively, our data indicate that OJS may be a safe drug although additional studies in the near future will be required before clinical trials can be taken. Drug safety is one of major issue in new drug development. The Food and Drug Administration (FDA) in most countries provides guidance on drug safety information for developing and disseminating new drugs with no or less toxicity as well as side effect. With the development of traditional medicine, concerns on the safety and potential toxicity of herbal medicines are increasing. However, there is little information of herbal medicine on the safety compared with synthetic drugs.wuji-san in China and goshaku-san in Japan) consisting of 17 medicinal herbs has long been used for treating common cold and illnesses including fatigue and gastrointestinal disorders in Asian countries including Korea, China and Japan. According to statistical data of the Korea National Health Insurance Corporation in 2011, OJS is the most frequently used herbal formulas of Oriental health treatments by 56 prescriptions. OJS is occupied 23% of the days of medication and 30% of medical expenses in Korea [Ojeok-san  rat model according to guidelines established by the Organization for Economic Cooperation and Development (OECD) for the testing of chemicals in accordance with the current regulations for Good Laboratory Practice Regulations . Our datThe OJS consisting of seventeen herbal medicines Table  and eachThe animal studies were conducted according to the guidance of the Institutional Animal Care and Use Committee in Korea Institute of Toxicology (KRICT)  under the Good Laboratory Practice Regulations for Non-clinical Laboratory Studies and approved by Korea Institute of Oriental Medicine Institutional Animal Care and Use Committee .Specific pathogen-free Crl:CD (SD) rats (n = 20/gender) were obtained from the Orient Bio Co.  and used after 2 weeks of quarantine and acclimatization. The animals were housed in a room maintained at 22 ± 3°C under a relative humidity of 50 ± 20% with artificial lighting from 08:00 to 20:00 and 12–15 air changes per hour. The animals were kept in stainless-steel wire-mesh cages and allowed sterilized tap water and commercial rodent chow  ad libitum.Healthy male and female rats were assigned to four groups  using Path/Tox System 4.2.2 . OJS extract was dissolved in distilled water for injection  and administered by oral gavage at doses of 0, 500, 1000 or 2000 mg/kg once a day for 4 weeks. Distilled water was given to the animals as the vehicle control. The daily dose (10 ml/kg body weight) of OJS was calculated based on the most recently recorded body weights of individual animals.Clinical signs and mortality were recorded twice a day (before and after treatment) throughout the study period. All clinical signs were recorded individually for type, observation day/time and duration using Path/Tox System 4.2.2 . The body weight of each rat was measured at the initiation of treatment and once a week during the study period. Food consumption was measured at the start of treatment and weekly throughout. Daily food consumption was determined by measuring the weight of chow supplied and remaining in metabolic cages each day. External eye examination was carried out during the last week of treatment with an indirect binocular ophthalmoscope , and the appearance of the conjunctiva, sclera, cornea, lens, and iris of each eye was recorded.During the last week of treatment, urinalysis was conducted on samples collected overnight in metabolic cages using a Multistix 10 SG  and urine chemical analyzer . Analysis included urine volume, glucose, bilirubin, ketone body, specific gravity, occult blood, pH, urobilinogen, and color.Animals were fasted overnight prior to blood collection or necropsy. Blood was drawn from the posterior vena cava under isoflurane anesthesia. Samples were collected in CBC bottles containing EDTA-2 K , and were analyzed to determine red blood cell count (RBC), white blood cell count (WBC), differential WBC count, hemoglobin concentration (HGC), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet (PLT) and reticulocyte (RET) using an ADVIA120 Hematology System (Bayer). Prothrombin time (PT) and activated partial thromboplastin time (APTT) were determined in blood samples treated with 3.2% sodium citrate using a coagulometer .For serum biochemistry, blood samples were centrifuged at 3000 rpm for 10 min and analyzed with an autoanalyzer . The analysis included alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (GGT), blood urea nitrogen (BUN), creatinine (CREA), creatine kinase (CK), glucose (GLU), total cholesterol (TCHO), albumin (ALB), albumin/globulin ratio (A/G), total protein (TP), triglyceride (TG), total bilirubin (TBIL), phospholipids (PL), sodium (Na), potassium (K), calcium (Ca), chloride (Cl), and inorganic phosphorus (IP).All surviving animals were anesthetized with isofluorane and sacrificed by aortic exsanguination prior to necropsy. Complete gross postmortem examinations were performed on all animals. Absolute organ weights were measured and relative organ weights (organ-to-body weight ratios) were calculated for the following organs: brain, pituitary gland, adrenal gland, liver, spleen, kidneys, heart, thymus, lung, salivary gland, thyroids, testes, ovaries, epididymides, seminal vesicle, prostate, and uterus.versus untreated control by following equation.Twenty-three of various cell lines were obtained from the American Type Culture Collection  or Korean Cell Line Bank . Cytotoxic effects of OJS against the cell lines were measured by 3--2,5-diphenyl tetrazolium bromide (MTT) assay as previously described . Cell vip < 0.05 or 0.01.Data collected during the study were examined for the variance homogeneity using Bartlett’s test. When Bartlett’s test indicated no significant deviation from the variance homogeneity, a one-way ANOVA was performed at α = 0.05. When significance was noted, a multiple comparison test (Dunnett’s test) was conducted to determine which pairs of groups were significantly different. Where significant deviations from variance homogeneity were observed, a nonparametric comparison test  was conducted. When a significant difference was observed in the Kruskal–Wallis test, the Dunn’s Rank Sum test was conducted to determine the specific pairs. Statistical analyses were performed using the Path/Tox System (ver. 4.2.2). The level of significance was taken as in vivo sub-acute toxicological analysis and in vitro cytotoxicity test.Recently, the use of herbal medicines is increased and thus the safety information is considered an important issue for herbal prescription. U.S. FDA has restricted the use of products containing aristolochic acid including botanical products marketed as traditional medicine since they can increase to occur in renal failure and renal cancer. It is of note that 11 kinds of herbal formulas are included in the restricted product list. Thus, safety report of medicinal herb or herbal formula is crucial step for their drug development and medication counseling. We previously reported that no mortality and no abnormality in clinical signs, body weight, and necropsy findings for any of the animals in the acute and sub-chronic toxicity study following oral administration of OJS, a Korean traditional herbal formula . In the Sub-acute toxicity of OJS extract was assessed in male or female Crl:CD (SD) rats (n = 24/gender) in accordance with OECD guideline. OJS extract was orally administered for 4 weeks at 500, 1000 or 2000 mg/kg/day. Body weight of male or female rats was increased in a time-dependent manner  of OJS extract for 24 h. As shown in Table in vitro biological studies. Further pharmacological studies will be required to find out the cause of WBC decrease by OJS in female rats before applying clinical studies in humans.We previously reported the safety of OJS in acute and sub-chronic toxicity as well as genotoxicity studies in rats. In addition, adverse effect of OJS has not been reported in clinical studies. Our current present study demonstrate that oral administration of Oriental herbal formula OJS causes leukopenia in female SD rats treated for 4 weeks. Thus, the no observed adverse effect level (NOAEL) values of OJS administration are determined to 2000 mg/kg/day in male and 500 mg/kg/day in female rats. Furthermore, no cytotoxic effect of OJS was found up to 200 μg/ml in 23 different cell lines. The cytotoxicity information of OJS can be used for


Allium tuberosum Rottler ex Spreng (ATRES) has been used as a traditional medicine for the treatment of abdominal pain, diarrhea, and asthma. In this study, we investigated the hair growth promoting activities of ATRES on telogenic C57BL6/N mice. Hair growth was significantly increased in the dorsal skin of ethanol extract of ATRES treated mouse group compared with the control mouse group. To enrich the hair promoting activity, an ethanol-insoluble fraction was further extracted in sequence with n-hexane, dichloromethane, ethyl acetate, n-butanol, and distilled water. Interestingly, we found that extraction with n-butanol is most efficient in producing the hair promoting activity. In addition, the soluble fraction of the n-butanol extract was further separated by silica gel chromatography and thin layer chromatography (TLC) resulting in isolating four single fractions which have hair growth regeneration potential. Furthermore, administration of ATRES extracts to dorsal skin area increased the number of hair follicles compared with control mouse group. Interestingly, administration of ATRES extract stimulated the expression of insulin-like growth factor-1 (IGF-1) but not of keratin growth factor (KGF) or vascular endothelial growth factor (VEGF). Taken together, these results suggest that ATRES possesses strong hair growth promoting potential which controls the expression of IGF-1. Hair plays an important role in body functions by protecting the head and maintaining the temperature of the head. Because of the increasing importance of hair in the field of beauty care, many researchers have focused on developing therapeutics for enhancing hair growth or preventing depilation . A uniqu Fructus panax ginseng extract has shown hair regeneration activity in C57BL6/N mice [ Eclipta alba extract was also reported to have hair growth promoting activity [ Polygonum multiflorum extract induces hair growth in resting hair follicles by upregulating Shh and beta catenin expression [Since the demand for novel therapeutics that provide safe hair growth enhancing results is growing, although most reports used the total extract of a herb to show the hair growth promoting activity without separating the substance into a single compound, many traditional herbs have been screened to determine whether or not they promote hair growth activity. It has been reported that Allium species known as Chinese chive that is widely distributed in East Asia. ATRES have been used for treating abdominal pain, diarrhea, hematemesis, and asthma in oriental medicine [ Allium tuberosum seeds extract was reported to have aphrodisiac properties [Allium cepa L.) to alopecia areata patients [ATRES is anmedicine . ATRES wmedicine . Allium operties  and, intpatients . HoweverATRES was obtained from a local Nonghyup market in Gangwon-do, Korea. Minoxidil was obtained from Hyundai Pharm. Co. Ltd.  and 1--3,5-diphenyltetrazolium bromide (MTT) was obtained from Sigma-Aldrich .5 cells/well were seeded in a 96-well plate and cultured for 24 h. The extracts were added to culture media and incubated at 37°C in CO2 incubator for 24 h, 48 h, and 72 h. MTT solution was then added and incubated for 2 h. Dimethyl sulfoxide  was used to dissolve the formazan crystals. The absorbance was measured at 540 nm with a microplate reader .Human hair dermal papilla cells (HHDPC) and human keratinocyte HaCaT cells were obtained from ScienCell Research Laboratories  and American Type Culture Collection , respectively. The cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM)  supplemented with 10% fetal bovine serum (FBS) , penicillin (100 U/mL), and streptomycin (100 ug/mL). Next, 1.5 × 10 in vivo experiments. The yield of ethanol extraction of ATRES was 19.5% . The insoluble fraction from the ethanol extract was then used for extraction by n-hexane and the insoluble fraction of the n-hexane extraction was used for the next extraction with dichloromethane. The insoluble fraction of dichloromethane extraction was used for the extraction with ethyl acetate and the insoluble fraction of ethyl acetate was then used for the extraction with n-butanol. Finally, the insoluble fraction of n-butanol was extracted with distilled water (DW). Each soluble fraction was concentrated, dried, and used for investigating the hair growth promoting activity. The yield of each dried extract was 3.27 g, 0.348 g, 0.21 g, 16.14 g, and 41.04 g, respectively.About 300 g of ATRES was extracted with 2000 mL ethanol at 40°C for 48 h. The extract was concentrated using a vacuum evaporator. It was then dissolved with 40% ethanol for subsequent treatment in the n-butanol soluble fraction was applied to the silica column and eluted with 400 mL of the methyl chloride/methanol (5 : 1) buffer. Each of the five fractions was tested using thin layer chromatography (TLC).About 700 g of C18 silica gel was packed into an open column . Three hundred milliliters of methyl chloride/methanol (5 : 1) buffer was preloaded into the column and then theEach of the five fractions from the silica gel chromatography was confirmed by thin layer chromatography (TLC). The TLC plate  was developed in chloroform/methanol/water (3 : 3 : 1) and it was visualized by heating at 80°C for 30 min after dipping in ethyl alcohol/sulfuric acid/anisaldehyde/acetic acid (90 : 5 : 1 : 1). After confirming the component, the fractionated samples, which contained the same component, were mixed, dried, and used for testing the hair growth promoting activity. ad libitum. For acclimation, all the mice were kept for one week before starting the experiment. These experiments were conducted in accordance with the Animal Research Committee of Sungkyunkwan University. The mice (n = 6 per group) were anesthetized with an intraperitoneal injection of a mixture of Rompun  and Zoletil . All the mice were shaved using animal clippers and further hair was eliminated using hair removal cream . To check the hair growth promoting activity, an ethanol extract of ATRES (3% or 5% in 150 uL of 40% ethanol) was applied topically to the dorsal back area every day for 14 days. The same volume of 40% ethanol and minoxidil  was used for the negative and positive controls, respectively. Soluble extracts (3% in 150 uL of 40% ethanol) of other solvents  were used for in vivo analysis.Six-week-old male C57BL6/N mice were obtained from Orient Bio  and housed in stainless steel cages under controlled temperature (23 ± 3°C), humidity (55 ± 10%), and photoperiod (12 h cycles of light and dark). All the animals were fed standard mice chow (Orient Bio) and water,2 incubator for 24 h. Total RNA was isolated with TRIzol reagent  according to the manufacturer's protocol. Changes in the expression levels of IGF-1, VEGF, and KGF were calculated using a real-time PCR machine  as previously described [The dorsal skin was isolated at the end of the animal study. HaCaT cell was treated with ethanol extract and incubated at 37°C in COescribed . The mRN2) was excised between the fore leg and the hind leg after being treated with ATRES extract. Each dorsal skin area was then fixed in 4% paraformaldehyde  and embedded in paraffin blocks. Next, 4 μm sections were stained with hematoxylin and eosin (H&E) and the representative area was photographed via light microscopy .The dorsal skin area , successively. Hair growth enhancing potential was tested by administering each of the soluble extracts to the telogenic mice. Although the soluble fraction of dichloromethane and ethyl acetate extraction showed no hair growth promoting activity in vivo (data not shown), the groups that were administrated n-butanol, n-hexane, and DW extracts showed enhanced hair growth 14 days after treatment , the hair growth promoting activity of n-butanol-n4 fraction was compared with the hair growth promoting activity of MXD, n-butanol, and the distilled water soluble fraction .Since theactivity , it was , and n4 , and the, and n4 . Since tfraction . As showd groups  in compa n-butanol, DW, and n-butanol-n4 fraction extracts by qRT-PCR ( n-butanol and distilled water showed increased levels of IGF-1 mRNA (P < 0.05) (P < 0.05)  . In addi < 0.05) . n-butanol-n4 fraction treated groups was significantly increased in comparison to the control group (P < 0.05), the n-butanol extract treated group showed the greatest increase in the number of hair follicles , but the hair growth promoting activity of each of the n-butanol extract subfractions was lower than that of the n-butanol extract (This study examined the hair growth promoting potential of ATRES through topical administration of extracts to the dorsal skin of C57BL6/N mice. ATRES is a well-known edible plant and it has been used in making traditional fermented food such as Kimchi in Korea. It was reported that consumption ofe stress  and consc cancer . Althougc cancer , 15, 16, in vivo . Recentl extract . Althoug n-butanol and DW extracts ( n-butanol extract, the mRNA expression of IGF-1 was significantly increased in the mouse group that was administered the n-butanol and DW extracts, in comparison to the control mouse groups ( n-butanol extract and the n-butanol and DW extracts ( in vivo and that the n-butanol extraction with silica gel chromatography method could be helpful way in developing a drug from ATRES. Further study should focus on identification, and functional studies of the four subfractions that were separated from the n-Butanol extract by silica gel chromatography should be conducted.Various growth factors have been proposed as controllers in hair growth. For example, VEGF, which is a well-known stimulator in angiogenesis, was reported to be a modulator in hair growth . Ginsenoextracts . Althouge groups . Interese groups . In addiextracts . These dThe present study shows that ATRES has strong hair promoting activity. We determined that butanol extraction is the most efficient way to enrich the hair promoting activity. In addition, four single components, which have hair growth promoting activity, were isolated by silica gel chromatography. Administration of ATRES extract to dorsal skin area increased the number of hair follicles and expression of IGF-1. Our study is the first attempt to investigate the hair promoting potential and its mechanism of action of ATRES extract.


Diabetes is characterized by chronic hyperglycemia and disturbances of carbohydrate, lipid and protein metabolism. We aimed to research association between serum lipid profile and blood glucose, hypothesizing that early detection and treatment of lipid abnormalities can minimize the risk for atherogenic cardiovascular disorder and cerebrovascular accident in patients with type 2 diabetes mellitus.Fasting blood glucose (FBG), total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TG) and glycated haemoglobin (HbA1c) levels were evaluated. A hepatic ultrasound was performed for every diabetic to evaluate hepatosteatosis. The study was done from January 2014 to June 2014 among 132 patients with T2DM who were admitted to outpatient clinic of Family Medicine department in a university hospital. The patients whose taking multi-vitamin supplementation or having hepatic, renal or metabolic bone disorders (including parathyroid related problems) were excluded from the study for the reason that those conditions might affect the carbohydrate and lipid metabolism in diabetes. Test of significance was calculated by unpaired student’s t test between cases and controls. Correlation studies (Pearson’s correlation) were performed between the variables of blood glucose and serum lipid profile. Significance was set at p<0.05.Results of serum lipid profile showed that the mean values for TC, TG, HDL and LDL in female patients were 227.6 ± 57.7 mg/dl, 221.6 ± 101.1 mg/dl, 31.5 ± 6.7 mg/dl and 136.5 ± 43.7 mg/dl, respectively. The mean values for TC, TG, HDL and LDL in male patients were 219.1 ± 34.7 mg/dl, 250.0 ± 100.7 mg/dl, 30.2 ± 7.4 mg/dl and 125.7 ± 21.4 mg/dl, respectively. Significantly higher mean serum levels of TC, TG and LDL and significantly lower mean serum levels of HDL were noted in patients with diabetes (p<0.001). FBG showed significant positive correlation with TC (p<0.05) and TG (p<0.05). Significant correlations were observed between serum levels of TC, TG, LDL and hepatosteatosis and HbA1c (p<0.05).The study showed widespread lipid abnormalities in the course of diabetes triggered dyslipidemia as hypercholesterolemia, hypertriglyceridemia, elevated LDL and decreased HDL. This study proposes the predominance of hyperlipidemia over increased prevalence of diabetic dyslipidemia. Diabetes mellitus is a common metabolic disorder characterized by absolute or relative deficiencies in insulin secretion and/or insulin action associated with chronic hyperglycemia and disturbances of carbohydrate, lipid and protein metabolism . SeveralIn the present study, we aimed to research association between serum lipid profile and blood glucose, hypothesizing that early detection and treatment of lipid abnormalities can minimize the risk for atherogenic cardiovascular disorder and cerebrovascular accident in patients with type 2 diabetes mellitus.2 (range: 26.10-44.98). Eleven out of 132 patients were obese in this study with BMI ranging from 31.02 to 44.98.We evaluated clinical and laboratory findings in 132 patients with T2DM. Of the total number of subjects, 61.4% (81) were females, and 38.6% (51) of the subjects were males . However, a significant difference was found as compared to normal range for related serum lipids of diabetics (p < 0.001) Table .Table 3Mean value of the TG/HDL ratio which is considered a surrogate marker of insulin resistance was 7.98 ± 3.87 among entire group. In correlation studies, TG/HDL ratio showed significant positive correlation with HbA1c (p < 0.01). Among all patients, hypercholesterolemia was found in 93 (70.5%) individuals. Similarly, increased LDL cholesterol was found in 117 (88.6%) individuals. Of the participants only 39 (29.5%) were found to have normal values for TG. Grade I and II hepatosteatosis were found in nearly one third of diabetics and Grade III hepatosteatosis in 63 47.7%) subjects while Grade IV hepatosteatosis was found in 27 (20.5%) of the study group by hepatic ultrasound  and TG (p < 0.05). The correlation of cholesterol with TG (p < 0.001) and LDL cholesterol (p < 0.001) were positive. However, HDL cholesterol showed a negative correlation with LDL cholesterol (p < 0.01). Findings were presented in Table  and TG  than males (38.6%) with T2DM in this study. The high proportion of females in this study may be due to the nature of population admitting to this hospital in that more of them seek medical attention than men under favour of having more free time because most of them were housewives.In patients with diabetes, many studies have clearly established that complications are mainly due to chronic hyperglycemia that exerts its injurious to health effects through several mechanisms: dyslipidemia, platelet activation, and altered endothelial metabolism –18. BothIn diabetes many factors may affect blood lipid levels, because of interrelationship between carbohydrates and lipid metabolism. Therefore, any disorder in carbohydrate metabolism leads to disorder in lipid metabolism and vice versa. Insulin resistance is a primary defect in the majority of patients with T2DM. In non-diabetic individuals insulin resistance in combination with hyperinsulinemia has a strong predictive value for future development for type 2 diabetes . SeveralAbnormal glucose reading is the commonest metabolic abnormality in people with T2DM accompanied by lower HDL levels, elevated LDL, hypercholesterolaemia, and hypertriglyceridaemia. Poor glycaemic control and hypertriglyceridaemia are significant biochemical abnormalities in patients with T2DM. Dyslipidemia management in people with diabetes mellitus, just like in any other individual, starts with a flawless evaluation that aims to identify secondary causes that might contribute to the abnormal lipid profile .The triglyceride-to-HDL cholesterol (TG/HDL) ratio has been investigated recently for various potential clinical uses in adult and paediatric populations , 32. PreAlthough obesity and T2DM commonly co-exist , mean BMLifestyle changes, including increased physical activity and dietary modifications, are the milestones of management. There is need for increased efforts by family members, community and healthcare professionals towards preventive based approach in the management of diabetes. This can be achieved through increased emphasis on lifestyle modification strategies such as exercise, increased dietary restrictions and weight control strategies especially for those with impaired fasting glucose. Monitoring of lipid profile by blood tests done in regular intervals in primary healthcare also might play an important role to detect and take care of lipid abnormalities both in diabetics and non-diabetics. One of the strengths of this study is that it was the first study in Istanbul, Turkey that looked at association between serum lipid profile and blood glucose among patients with T2DM in primary healthcare. To the best of our knowledge, there has not been any previous study both performing blood tests and hepatic ultrasound on this group of subjects. Despite several important findings in the present study, relatively small sample size is considered as a limitation of it.The highest priority for diabetic individuals who have poor glycaemic control should be to achieve almost near normal blood glucose levels, with the expectation that this effort will also cause an improvement in dyslipidemia , 36, 37.The present study suggested that common lipid abnormalities during diabetes induced dyslipidemia are hypercholesterolemia, hypertriglyceridemia and elevated LDL cholesterol. Results suggest a high prevalence of dyslipidemia, which might be playing a major role in the development of cardiovascular diseases and cerebrovascular accidents among diabetic patients. The optimal care for diabetic patients should include routine monitoring of blood glucose and serum lipid profile. Efforts to achieve lifestyle changes, such as weight reduction, physical exercise and smoking cessation should be encouraged and initiated first and then followed by medication with lipid lowering drugs prescribed in evidence-based necessary conditions. The optimum treatment with proper antidiabetic drugs to obtain fair glycaemic control should go concomitantly with lipid lowering drugs as well as with taken dietary precautions. When those mentioned above are done, we might properly speak about the benefits that primary healthcare could ensure to diabetics by the advantage of monitoring patients more closely.The cross-sectional study sample included Turkish participants diagnosed with T2DM. The study was done from January 2014 to June 2014 in the outpatient departments of Family Medicine clinic of the Bezmialem Vakif University Hospital in the largest city of Turkey, namely Istanbul, in the north-west region of the country and participating 132 subjects with T2DM (more than 6 months duration) were randomly selected and recruited in the study. The patients whose taking multi-vitamin supplementation or having hepatic, renal or metabolic bone disorders (including parathyroid related problems) were excluded from the study for the reason that those conditions might affect the carbohydrate and lipid metabolism in diabetes , 39. AlsSubjects were requested to visit the family medicine outpatient clinic following an overnight fasted state (≥8 hours) for anthropometry and blood withdrawal by the clinic nurse and physician on duty, respectively. Anthropometry included height (rounded to the nearest 0.5 cm), weight (rounded to the nearest 0.1 kg), and mean systolic and diastolic blood pressure (mmHg). Body mass index (BMI) was calculated as weight in kilograms divided by height in square meters. Fasting blood samples were collected and transferred immediately to a non-heparinized tube for centrifugation. Collected serum was then transferred to pre-labeled plain tubes and delivered to the bio-chemistry laboratory in Bezmialem Vakif University Hospital.Fasting glucose, cholesterol and triglyceride were measured using an chemical auto-analyzer . Cholesterol esters will be hydrolyzed by cholesterol esterase. Cholesterol will be oxidized into cholest-4-en-3-on and hydrogen peroxide by bacterial cholesterol oxidase. Hydrogen peroxide in the presence of phenol and amino-4-antipyrine forms a complex of red color showing absorption maximum between 500–550 nm. It is based on the principle that triglycerides in the serum sample are hydrolyzed enzymatically by the action of lipase to glycerol and fatty acids. The glycerol formed is converted to glycerol phosphate by glycerol kinase (GK). Glycerol phosphate is then oxidized to dihydroxyacetone phosphate by glycerol phosphate oxidase (GPO). The librated hydrogen peroxide is detected by a chromogenic acceptor, chlorophenol-4-aminoantipyrine, in the presence of peroxidase (POD). The red quinone formed is proportional to the amount of triglycerides (TG) present in the sample and is measured at 546 nm. High density lipoprotein (HDL) is measured by using phosphotungstate precipitation method based on the principle that chylomicrons, very low density lipoprotein (VLDL) and low density lipoprotein (LDL) fractions in serum or plasma are separated from HDL by precipitating with phosphotungstic acid and magnesium chloride. After centrifugation the cholesterol in the HDL fraction which remains in the supernatant is assayed with enzymatic cholesterol method using cholesterol esterase, cholesterol oxidase, peroxidase and the chromogen 4-aminoantipyrine. The LDL cholesterol test is a two reagent homogenous system. The assay is comprised of two distinct phases. In phase one a unique detergent solubilizes cholesterol from non-LDL-lipoprotein particles. This cholesterol is consumed by cholesterol esterase, cholesterol oxidase, peroxidase and 4-aminoantipyrine to generate a colorless end product. In phase two a second detergent in reagent 2 releases cholesterol from the LDL lipoproteins. This cholesterol reacts with cholesterol esterase, cholesterol oxidase and a chromogen system to yield a blue color complex which can be measured bichromatically at 540/660 nm. The resulting increase in absorbance is directly proportional to the LDL concentration in the sample. The value of VLDL cholesterol was calculated as one-fifth of the concentration of triglycerides.It should be noted that the Quality Assurance (QA) standards are maintained by TS EN ISO 15189, whereas the QA department audits the laboratory at regular intervals. Ethical approval was taken from the institutional review board (IRB) prior to the commencement of the study.Ultrasound could be used contributively in detecting hepatosteatosis and it is a non-invasive and easily applicable method. However, liver biopsy remains the only accurate way to diagnose hepatosteatosis. There are 4 sonographic findings of diffuse fatty change in the liver characterising different grades: (1) a diffuse hyperechoic echotexture (bright liver), (2) increased liver echotexture compared with the kidneys, (3) vascular blurring, and (4) deep attenuation . A hepatData were analyzed using the Statistical Package for the Social Sciences version 16.0 . Normal continuous variables were presented as mean ± standard deviation. Test of significance was calculated by unpaired student’s t test between cases and controls. Correlation studies (Pearson’s correlation) were performed between the variables of blood glucose and serum lipid profile. Significance was set at p < 0.05.


Background. Hepatic angiosarcoma is a rare and aggressive tumor that often presents at an advanced stage with nonspecific symptoms. Objective. To report a case of primary hepatic angiosarcoma in an otherwise healthy man with normal liver function tests two months prior to presenting with a short period of jaundice that progressed to fulminant hepatic failure. Methods. Case report and review of literature. Conclusion. This case illustrates the rapidity of progression to death after the onset of symptoms in a patient with hepatic angiosarcoma. Research on early diagnostic strategies and newer therapies are needed to improve prognosis in this rare and poorly understood malignancy with limited treatment options. A 75-year-old man presented to our hospital with three-week history of anorexia, abdominal fullness, chills, and three-day history of jaundice. He was diagnosed with AV nodal reentry tachycardia (AVNRT) six months earlier which terminated with the Valsalva maneuver. His medication included Diltiazem and Aspirin. He had no further episodes of AVNRT and liver enzymes were normal at routine follow-up in the cardiology clinic two months before presentation. On admission he was afebrile and reported no abdominal pain, hematemesis, melena, or pale stools. Physical examination revealed a frail looking man with skin and scleral icterus, normal heart sounds, and blood pressure. Abdominal examination revealed epigastric tenderness without organomegaly or ascites. Laboratory tests were significant for total bilirubin 9 mg/dL, albumin 4.0 g/dL, alkaline phosphatase 383 U/L, gamma-glutamyl transferase 701 U/L, aspartate aminotransferase, 104 U/L, alanine aminotransferase 206 U/L, and lactate dehydrogenase 451 U/L. Carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP) levels were normal and CA19-9 was 38.8 IU/mL. Human immunodeficiency virus (HIV) and hepatitis panel were negative. Chest X-ray was normal and ECG showed sinus tachycardia. Abdominal ultrasound scan was negative for cholelithiasis and cholecystitis. Contrast enhanced triple-phase computerized tomography scan of his abdomen showed a hepatic mass measuring 13 × 11 × 10 cm with small dilated bile ducts surrounding the mass . There wHepatic angiosarcoma (HAS) is a very rare and aggressive tumor that accounts for about 1.8–2% of all primary liver cancers. Epidemiologically, it is associated with exposure to polyvinyl chloride (PVC), thorium dioxide (thorothrast), arsenic, and inorganic copper , 4. BaseThe presenting symptoms of hepatic angiosarcoma are nonspecific and may be confused with any other hepatobiliary disorder . The mosFulminant liver failure (FHF) due to HAS is very rare and the mechanism of liver failure is multifactorial. There have been six cases of FHF secondary to HAS reported in the literature to date and the available evidence suggests that a combination of hepatic ischemia leading to parenchymal infarction, vascular occlusion of portal vein flow by tumor, thrombi, and nonocclusive infarction of the liver due to shock from secondary causes such as sepsis or cardiac dysfunction may play a role –19. DissTreatment of the tumor to date is empirical . Radical


Metacyclogenesis was evaluated by different criteria, namely, promastigote size , surface modifications  binding, complement resistance), and infectivity to human macrophages. Growth curves were similar for all parasites evaluated. The various features analyzed were expressed in a high percentage of promastigotes at 6th and 10th days of culture and a low percentage at the 2nd day. However, in most isolates, these features, considered as markers of metacyclogenesis, seemed to develop with different time courses, since the percentages of metacyclic forms detected with each technique were usually different. Parasites from 6th or 10th day and those negatively selected with lectin or mAb similarly infected human macrophages. From all isolates analyzed, L. amazonensis PH8 and MAB6 showed the highest and the lowest levels of susceptibility, respectively, to leishmanicidal activity of IFN-γ/LPS-activated macrophages. Our results showed that by using different techniques to evaluate different aspects of metacyclogenesis  different percentages of metacyclic promastigotes can be detected in each isolate culture.This study was designed to assess  Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis which lead to localized cutaneous leishmaniasis (LCL) but also can cause severe clinical forms as mucosal (ML) and diffuse cutaneous leishmaniasis (DCL), respectively [American tegumentary leishmaniasis (ATL) is mainly caused byectively –3. The oInfection of hosts occurs by the injection of infective forms, known as metacyclic promastigotes, by parasite-harboring sand flies during blood meal. Metacyclic promastigotes develop inside the insect vector from procyclic promastigotes, less infective, which are newly transformed from amastigotes sucked with infected host cells. Metacyclogenesis can also occur in axenic cultures of parasites from the logarithmic to the stationary phase of growth –6. Along L. major [ Leishmania [ L. braziliensis and L. amazonensis [ Leishmania, most studies evaluating infectivity of New World Leishmania promastigotes forms have used murine macrophages [Although metacyclogenesis has been well studied inL. major –7, 19 anishmania , 20, 21,rophages , 21, 22,rophages –25. More in vitro metacyclogenesis of L. (V.) braziliensis and L. (L.) amazonensis clinical field isolates by parasite morphology, surface changes, and infectivity to human macrophages, comparing the results with those obtained from the well-studied Leishmania reference strains .In the present study, we evaluated the L. braziliensis (MHOM/BR/1975/M2903), named herein as M2903, and L. amazonensis (IFLA/BR/67/PH8), named herein as PH8, were used. The clinical field isolates were obtained from ATL patients with different clinical forms  and characterized as L. braziliensis and L. amazonensis, as we previously described [ L. braziliensis, MHOM/BR/2003/IMG (IMG3), obtained from LCL lesion, and MHOM/BR/2006/PPS (PPS6m), obtained from ML lesion, and one isolate of L. amazonensis MHOM/BR/2006/MAB (MAB6), obtained from DCL lesion, were assessed.Known strains ofescribed –28. Two μg/mL streptomycin (Sigma), at 26°C, pH 5.8. Parasites were maintained in logarithmic phase of growth by passages each 2-3 days, always starting the cultures with 5 × 105 parasites/mL. Only 3rd–6th passage parasites were used in the experiments.Promastigotes were all stored in logarithmic phase of growth (2-3 days of culture) in liquid nitrogen , after differentiation from amastigotes derived from lesions of interferon gamma-knockout mice . CulturePromastigotes growth curves were obtained by quantification of parasites after dilution of a culture aliquot in phosphate-buffered saline (PBS) containing 0.3% paraformaldehyde, in hemocytometer, during 12 days.After 2, 6, or 10 day of culture, parasites were harvested and washed with PBS . The pellet was suspended in 5% FCS-containing PBS before centrifugation onto slides . After staining , photomicrographs were taken under a light microscope . The images were analyzed using the software Image J . The cell body size and flagellum lengths were measured for 300 parasites in each day, and the percentage of metacyclic forms was calculated  were incubated with 100 μg/mL Bauhinia purpurea lectin , which binds to galactosyl  N-acetylgalactosamine -GalNAc) present in LPG of procyclic promastigotes or 100 μg/mL 3A1-La mAb, which binds to LPG of procyclic promastigotes, but not of metacyclic forms [A previously described technique to identify and select the metacyclic promastigotes  was usedic forms , 18, 29,μL) for flow cytometry analyses. Two thousand events were acquired using a FACScan Flow Cytometer  and parasites were analyzed after plotting the FSC × SSC [high) and metacyclic (FSClow) forms.Total parasites from 2nd, 6th, or 10th day of culture or negatively selected metacyclic forms (with BPL or 3A1-La mAb) were washed with PBS , quantified, and fixed with 1% paraformaldehyde in PBS (300 SC × SSC  using th5) were incubated with 10% rabbit serum at 37°C, for 1 h. The suspension was kept at 4°C while quantification of live and dead parasites was carried out under a light microscope. Results were expressed as percentage of survived parasites, the complement resistant forms, in comparison with nontreated parasites.Lyophilized rabbit serum  was reconstituted in Hanks Balanced Salt Solution  and added to parasites at the concentration of 10% . Parasites from 2nd, 6th, or 10th day of culture were collected and washed with HBSS . Parasites suspended in HBSS ), were distributed onto 13 mm-round glass coverslips  and placed in 24-well plates . Culture medium was replaced each 48 h during 7 days . Nonadherent cells were removed during medium changes and adherent monocytes differentiated into macrophages. For some experiments, at the 6th day of culture cells were treated with 10 ng/mL of recombinant human IFNγ (Sigma) overnight, before macrophage infection.All procedures for macrophage derivation were approved by local Ethical Committee . Venous blood samples (~12 mL) were obtained from healthy blood donors  in EDTA-vacuum tubes . Blood was layered on Ficoll gradient  and centrifuged at 1,000 g, r.t., 20 min. Mononuclear cells (5 × 105 parasites/well) at a multiplicity of infection (MOI) ~10 : 1 (considering that monocytes comprise ~10% of the mononuclear cells). Cultures preincubated with recombinant human IFNγ  were treated with lipopolysaccharide  at the same time of parasite addition. Cultures were incubated for 24 h or 72 h, 36°C, 5% CO2. After incubation, cells were fixed and stained with commercial Instant Prov kit (Newprov) and analyzed under a light microscope  to determine the infection index. Three to five hundred cells were analyzed in random fields and the percentage of infected cells and the mean number of intracellular amastigotes per infected cell (at least 50 infected cells were counted) were determined. Infection index = percentage of infected cells × mean number of parasites per infected cell. All assays were done in duplicate and cell infection was blinded evaluated.Growth stationary phase promastigotes or metacyclic forms isolated by negative selection with BPL/3A1-La mAb were washed with PBS . Parasites were added to macrophage cultures .For each evaluation, 3 to 6 experiments in triplicates were performed. Data are shown as median with interquartile range (minimum and maximum) or mean ± EPM. Mann-Whitney, Wilcoxon matched pair or http://dx.doi.org/10.1155/2014/393049). Next, we evaluated the morphological alterations to identify metacyclic forms in cultures.  L. amazonensis isolates present larger cell body and lengthier flagellum than L. braziliensis isolates  were detected ( L. amazonensis (PH8 and MAB6)   were couhigh) and metacyclic (FSClow) forms. L. braziliensis) and MAB6 (L. amazonensis) isolates, at the 2nd  increased along culture time. High frequencies of smaller forms were detected at 6th and 10th days of cultures of both L. braziliensis -GalNAc present in LPG of L. braziliensis procyclic promastigotes, and the mAb 3A1-La, which binds to L. amazonensis procyclic forms, was used here to identify and try to purify the metacyclic promastigotes. Similarly to what we found using the morphology criterion, a significant increase of nonbinding forms from L. braziliensis and L. amazonensis isolates occurred with the negative selection with BPL .As expected, the nonagglutinated promastigotes isolated by negative selection were mostly the small forms as seen by the enrichment in FSC L. major and L. amazonensis [ L. braziliensis and L. amazonensis change in LPG composition during in vitro metacyclogenesis. Thus, parasite isolates were also assayed for complement resistance to compare the percentage of resistant forms in the cultures . Likewise, we observed that the resistant forms were much more frequent at the stationary phase of parasite growth (6th and 10th days) than at logarithmic phase (2nd day) .γ, then LPS addition). Similar to that observed with nonactivated macrophages . In parallel, known strains of L. braziliensis (M2903) and L. amazonensis (PH8) were also studied. Our results showed a similar growth profile among all isolates , which was also similar to the one previously described for M2903 strain, even when different culture media were used [ in vitro passages. It has been shown that L. braziliensis and L. amazonensis in cultures can present similar growth depending on the medium components [ L. braziliensis and L. amazonensis have been described [In the present study we evaluated theere used , 32. Sinere used , the parescribed . In the  L. braziliensis [ L. amazonensis [ L. amazonensis metacyclic forms are larger than L. braziliensis (6th and 10th days of culture)  but different for L. amazonensis (Josefa strain), for which we found higher percentages of metacyclic forms. In that case, since the authors evaluated the parasites at 5th day of culture it is possible that a difference of 24 h could be enough to increase the percentage of metacyclic forms detected in the present study (6th day). Although morphometric and cytometric analysis are based on morphological alterations, these techniques detected different percentages of small parasites forms (metacyclic forms) in L. amazonensis and also in L. braziliensis cultures. It is clear from our experiments that cultures of L. braziliensis present more clusters of parasites than L. amazonensis which can be detected as large forms in flow cytometry. In addition, L. amazonensis presents less alteration of body size than L. braziliensis  and MAB6 (L. amazonensis) isolates the percentages of complement-resistant and BPL- or mAb-nonagglutinated forms did not match at 6th-day cultures  it can be suggested that complement resistance can be acquired hours/days after carbohydrate alterations, suggesting that other factors/molecules not analyzed here are involved.Another characteristic of metacyclic forms, also related to LPG, gp63 and probably other protein modifications, is the increase of parasite resistance to the complement system , 13, 14.cultures . These r in vitro metacyclogenesis of different clinical field isolates of Leishmania, besides understanding the parasite biology, is to establish optimal conditions to investigate the infection by Leishmania, particularly the human macrophage infection, since most studies are carried out using the murine model. Our results showed that parasites from 6th or 10th day of culture were more efficient to infect macrophages than promastigotes forms from 2nd day (L. braziliensis) there was a significant decrease in infection index from 24 h to 72 h of incubation when macrophages were infected with parasites from the 2nd or 10th day of culture. These results suggested that 2nd-day promastigotes could not be well adapted to survive in macrophages and those from 10th day are weakened because of nutrient starvation. In both cases parasites were more susceptible to microbicidal activity of macrophages. One possibility is that parasites from the 2nd day activate macrophages during the first hours of infection inducing microbicidal mechanisms whereas those from 6th day silently infect macrophages. Ueno et al. [ Leishmania parasites from logarithmic phase of growth, instead of metacyclic forms, use mannose receptor during macrophage phagocytosis activating parasite killing mechanisms. Macrophages infected with 6th-day parasites did not present significant increase of the infection index until 72 h, suggesting that human macrophages allow the infection but do not favor parasite growth. Likewise, Hsiao et al. [ L. infantum chagasi until 72 h after in vitro infection with metacyclic forms. Thus, our results suggest that the infection with L. braziliensis or L. amazonensis can be controlled in human macrophages during 72 h. Although the mechanisms of parasite control need to be investigated, it has been shown that reactive oxygen species (ROS) are responsible for controlling New World Leishmania infection in human macrophages [One important reason to study 2nd day . These ro et al.  have shoo et al.  have demrophages , 39. in vitro differentiation process it can be not possible to detect differences in the outcome of human macrophage infection with stationary phase parasites or selected metacyclic forms of New World species of Leishmania. Pinto-Da-Silva et al. [ L. braziliensis (cultures with 50% of metacyclic forms detect by this technique) were more infective to murine macrophages than parasites from stationary phase of growth, but without statistical significance. Together, these results suggest that depending on the number of metacyclic forms present in the parasite preparations and the degree of parasite virulence, to enrich metacyclic forms could be need to obtain high levels of macrophage infection. Still, infection with metacyclic forms enriched by negative selection can induce more homogenous results when evaluating the quality of immune responses during L. braziliensis or L. amazonensis human macrophage infection.In the present study, negative selection by agglutination assay was used to obtain selected metacyclic forms and to compare infection of human macrophages with parasites in stationary phase of growth  or metacyclic forms . No significant differences were found in macrophage infection index with total parasites or selected metacyclic forms, either after 24 h or 72 h. This result confirms that the majority of promastigotes on the 6th day of culture is metacyclic, which is also strengthened by the other features analyzed here. Also, after 6 days of culture low amounts of parasites can be procyclic forms and parasites that are differentiating into metacyclic forms could be able to infect macrophages as well as metacyclic forms. Moreover, the isolation of metacyclic forms by negative selection does not ensure 100% efficiency and it is possible that the process only enriches the population in metacyclic forms, so that a low percentage of procyclic forms may be present in the lectin or mAb nonagglutinated population. Thus, due to a continuousa et al.  showed tγ plus LPS; [ Leishmania species [γ/LPS. This treatment reduced the infection index caused by PH8, but not for other isolates . Cl species , 42–44. isolates , suggest in vitro metacyclogenesis of L. braziliensis and L. amazonensis clinical field isolates using, simultaneously, four well established techniques. Although the metacyclogenesis profile of all isolates was similar, depending on the technique used different density of parasites with features of metacyclic forms were detected, indicating that the different features evaluated do not develop simultaneously. Whereas for both species negative selection assay (with BPL or 3A1-La mAb) was the technique that identified higher percentages of parasites with biochemical characteristics of metacyclic promastigotes, morphometry could identify high amounts of L. braziliensis small promastigotes, but not of L. amazonensis. On the other hand, flow cytometry detected lower percentage of L. braziliensis small forms, but high amounts of L. amazonensis. Morphometric and cytometric analysis although based on morphological aspects did not detect the same amounts of metacyclic forms , as well as complement resistance and negative selection assays, both detecting LPG alterations, were not in accordance for PPS6m and MAB6 isolates. Our results showed that by using different techniques to evaluate different aspects of metacyclogenesis  different percentages of metacyclic promastigotes can be detected in each isolate culture. These results suggest that a continuous differentiation process from procyclic into metacyclic forms is working in vitro providing different stages of parasite differentiation.This is the first study onThus, according to our results, for each new clinical field isolate to be studied is important to determine the metacyclogenesis profile by different techniques, and to identify the better day of parasite culture to select metacyclic forms to infect macrophages, especially in the case of low rates of metacyclogenesis. To use the same amounts of metacyclic forms to compare the ability of different parasites to infect macrophages, especially when parasite cultures present huge differences in percentage of metacyclic forms, can be determinant in virulence research. L. braziliensis and L. amazonensis although showing similar time course of metacyclogenesis (peak of metacyclic forms at 6th day of culture) can display different abilities in establishing a successful infection in nonactivated or activated human macrophages. The results suggest that in vitro metacyclogenesis is a continuous differentiation process from procyclic into metacyclic forms. Thus, the different features acquired by promastigotes during in vitro culture , which are used as criteria to evaluate metacyclogenesis, may develop with different time courses.In our study, clinical field isolates ofL. braziliensis) and MAB6 (L. amazonensis) as well as M2903 (L. braziliensis) and PH8 (L. amazonensis) strains were incubated into Grace's insect medium, at 26°C. Parasites were quantified in hemocytometer every day for 12 days. Figure 1 shows that all parasites presented a similar time course profile of in vitro growth.To stablish the time course profile of all isolates evaluated in the present study, clinical field isolates IMG3, PPS6m  and L. amazonensis (MAB6 and PH8) in vitro cultures. The morphometry of fixed and stained parasites was done by using the software Image J. As showed in Table 1, length of the parasite cell body decreased whereas length of flagellum increased along the culture time (6th and 10h days of cultures) for all isolates, indicating the differentiation of procyclic (more frequent at the 2nd day of cultures) into metacycic forms.The parasite morphometric analysis was performed in the 2nd, 6th or 10th day after starting To compare the methods used to evaluate metacyclogenesis using different criteria, all results were showed in Table 2: morphometry, cytometry, negative selection of metacyclic forms with lectin/mAb and complement resistance assay. Results showed that different features to evaluate metacyclogenesis may develop with different time courses. In this study, this was especially observed with PPS6m isolate.It is known that metacyclic forms are more infective than procyclic forms, therefore human monocyte-derived macrophages were incubated with non-selected  or selected (non-agglutinated) parasites  from 6th day of culture. The infection index was evaluated after 24 h or 72 h of incubation. Results showed that for those isolates/strains it is not necessary to enrich the parasite suspension in metacyclic forms as the data showed a similarity between infection index using total or selected parasites.


Cyclin D1 (CCND1) is critical in the transition of the cell cycle from the G1 to S phases, and unbalanced cell cycle regulation is a hallmark of carcinogenesis. Numerous epidemiological studies have evaluated the association between the CCND1 A870G polymorphism and the risk of prostate cancer (PCa). However, these studies have yielded conflicting results.In the present study, the possible association above was assessed by a meta-analysis. Eligible articles were identified for the period up to July 2014. Pooled odds ratios (ORs) with 95% confidence intervals (95% CI) were appropriately derived from fixed effects or random effects models.P = 0.380; OR = 1.088, 95% CI = 0.896 to 1.321 for AA versus GG, P = 0.393; OR = 1.044, 95% CI = 0.941 to 1.158 for GA versus GG, P = 0.414; OR = 1.053, 95% CI = 0.955 to 1.161 for the dominant model AA + GA versus GG, P = 0.303; OR = 1.072, 95% CI = 0.881 to 1.306 for the recessive model AA versus AA + GA, P = 0.486). Moreover, subgroup analyses according to ethnicity failed to demonstrate a significant association between this polymorphism and prostate cancer. In addition, we also performed a stratified analysis of cases with PCa metastasis, and the results supported the findings of no significant association between CCND1 A870G polymorphism and metastasis risk of PCa.A total of ten case-control studies, which included 3,820 cases and 3,825 controls, were identified. Overall, the allelic/genotypic association between the G870A polymorphism and prostate cancer was nonsignificant (OR = 1.045, 95% CI = 0.947 to 1.153 for A versus G, Our results suggest that the CCND1 A870G polymorphism might not be a potential candidate for predicting prostate cancer risk, including metastasis risk. As one of the most frequent malignant diseases among men, prostate cancer (PCa) is a complex disease that is caused by a multitude of factors . RecentlCyclin D1 mRNA is alternatively spliced to transcribe two different transcripts, which yield to functionally different proteins [CCND1 is involved, alternatively, in splicing of CCND mRNA [Cell cycle dysregulation plays a critical role in a variety of malignancies and contributes to an increased risk of metastasis, in part, by altering the cell’s ability to respond appropriately to DNA damage ,5. A numproteins ,16, and CND mRNA .CCND1 A/A genotype and the risk of various cancers. Qin et al. found that the CCND1 A/A genotype may increase brain tumor risk, especially for gliomas [et al. also reported that an A870G polymorphism in CCND1 confers an increased risk for breast cancer [et al. revealed that a CCND1 G870A polymorphism has no association with esophageal cancer risk in ethnicity and histology [CCND1 A870G polymorphism and prostate cancer susceptibility, the conclusions have been inconsistent. Wang et al. reported that the A allele of the CCND1 A870G polymorphism was recessively associated with susceptibility to PCa [CCND1 A870G polymorphism and prostate cancer was found by Chen et al. [Epidemiological studies have reported the association between the  gliomas . Yang ett cancer . Howeveristology . Althougy to PCa . But no n et al. .CCND1 A870G polymorphism and prostate cancer risk, which were previously published, were analyzed via a meta-analysis to examine a more specific association between the CCND1 A870G polymorphism and prostate cancer risk and various published observational studies.In the present study, ten case-control studies on the CCND1/cyclin D1, prostate cancer/carcinoma/tumor, variant/genotype/polymorphism/SNP, and the combined phrases for all genetic studies on the association between the CCND1 A870G polymorphism and prostate cancer risk. We also checked the references of the retrieved studies and reviews to ensure the complement of this meta-analysis. This work was approved by the ethics committees of Zhejiang Provincial People’s Hospital.The PubMed, MEDLIN, and Web of Science databases were searched for studies published before July 2014. The key words used for searching were as follows: CCND1 polymorphism and prostate cancer risk, and (c) sufficient published data for both patients and controls.The following inclusion criteria were used for the literature selection in our meta-analysis: (a) case-control study, (b) evaluation of the The following exclusion criteria were set: (1) incomplete raw data, (2) repetitive reports , (3) materials and methods were not well-described and reliable, or (4) not an English paper.Two investigators extracted information independently according to the inclusion and exclusion criteria listed. When it came to conflicting evaluations, an agreement was reached after a discussion. For each eligible study, we collected information as follows: the first author’s name, publication data, country of origin, sources of controls, number of different genotypes, and ethnicity of the study population (categorized as Asian and Caucasian). For studies including subjects of different ethnic groups, data were extracted separately for each ethnic group whenever possible.χ2 of Fisher’s exact test). Heterogeneity and cumulative analysis were assessed by χ2-based Q test. OR estimation was calculated with the fixed effect model (Mantel-Haenszel method) when statistical heterogeneity did not exist (P > 0.10). Otherwise, the random effects model (DerSimonian and Laird method) was selected. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of association between the CCND1 polymorphism and prostate cancer risk. The pooled ORs were performed for a codominant model (GA versus GG and AA versus GG), dominant model (GA/AA versus GG), and recessive model (AA versus GA/GG). Stratified analyses were also performed by ethnicity. Publication bias was evaluated by Funnel plots, and Begg’s test and Egger’s test were also used to detect publication bias. Statistical analysis was performed using STATA versions 11.0 .The Hardy-Weinberg equilibrium (HWE) for the control group of each study was assessed using a goodness-of-fit test (P > 0.05) in studies except for Mandal et al. (P = 0.013) and Koike et al. (P = 0.004).After an extensive search, a total of 43 relevant studies were identified Figure . FollowiCCND1 A870G polymorphism and prostate cancer risk. The combined results based on all studies revealed that variant genotypes are not associated with increased prostate cancer risk in different genetic models  , and the random effects model was employed in these studies. We did not find significant heterogeneity for the following model , and a fixed effects model was performed.The following genetic model of the ten studies showed statistically significant heterogeneity using the CCND1 G720A polymorphism . This result was confirmed by the Begg’s rank correlation method  , which increased the frequency of alternative splicing and encoded a protein with an altered C-terminal domain and increased the stability or half-life of the protein, has garnered wide attention. To date, several studies have reported the role of the CCND1 G870A polymorphism in prostate cancer risk [CCND1 G870A polymorphism associated with prostate cancer, we conducted a meta-analysis of results from ten case-control studies on the association of the CCND1 G870A polymorphism with PCa risk. Overall, a total of 3,820 cases and 3,825 controls were included. However, the results indicated that no significant association between the CCND1 A870G polymorphism and PCa risk was found.Cell cycle regulation plays an important role in the evolution of cancer by influencing cell proliferation, differentiation, and apoptosis . CCND1, cer risk ,22,24. Bet al. [CCND1 A allele was more frequently observed in the PCa group than the control group, and men with the AA genotype had an increased risk of PCa compared to those with the GG genotype. Mandal et al. [CCND1 AA genotype was observed to be associated with a significant increase in PCa risk. However, Koike et al. found that no significant association of the genotype frequency of the CCND1 with overall cases and controls [CCND1 A870G polymorphism and PCa risk was found.Wang et al.  showed tl et al.  also revcontrols . Our preCCND1 A allele showed significantly increased risk of PCa metastasis [et al. did not find any significant risk when analyzing data for the risk of susceptibility for metastasis with the CCND1 polymorphism [CCND1 A870G polymorphism and metastasis risk of PCa, as reported by Mandal et al. [et al. [CCND1 A870G polymorphism might be in close linkage with different nearby causal variants in one ethnic population but not in another, according to the report of Yu et al. [Stratification analysis showed that the tastasis . But Manmorphism . In our l et al. . But the [et al. . Base on [et al. . Differeu et al. . In addiCCND1 A870G polymorphism and did not evaluate other genes or polymorphisms; whether this polymorphism integrated with other risk factors to enhance the predictive power requires further study.However, some limitations of this meta-analysis should be addressed. First, the results were based on the unadjusted estimates with original data from these collected studies being unavailable, which limited the evaluation with certain covariates, including cancer type, smoking, drinking, age, and other environmental factors. Second, the controls of several studies were various in the analysis, which may have induced the bias of the results and prevented the drawing of more detailed conclusions. Third, the single-locus-based nature of this meta-analysis precluded the possibility of gene-gene and gene-environment interactions; the analysis did not consider these factors because of the lack of sufficient data. Furthermore, we only concentrated on the CCND1 A870G and prostate cancer susceptibility. In addition, our observations raise the question of a potential heterogeneous effect of A870G across different ethnic populations. Nevertheless, for practical reasons, we hope that additional studies of CCND1 that include functional DNA repair gene polymorphisms in a large cohort of different ethnicities, combined with more appropriate methods, will augment the etiology of the pathogenesis of PCa.In conclusion, we expanded previous individually underpowered studies and suggested that no obvious association was found between the