Impact of sarcopenia and low muscle attenuation on outcomes of ovarian cancer: a systematic review and meta-analysis.

OBJECTIVE: The aim of this study was to examine the association of sarcopenia and low muscle attenuation with survival and other clinical outcomes in patients with ovarian cancer. MATERIALS AND METHODS: Systematic search was done in PubMed, EMBASE and Scopus databases for observational studies that documented the link between sarcopenia and outcomes of interest in patients with ovarian cancer, with long-term survival as a primary outcome. Other outcomes included risk of recurrence, progression-free survival and complications. Pooled effect sizes were reported as hazards ratio (HR), relative risk ratio (RR) or weighted mean difference (WMD). Random effects model was used for the analysis. RESULTS: Twenty-two studies were selected, of which all, except one, were retrospective in design. Low skeletal muscle index (SMI, indicating muscle mass) (HR 1.30, 95% CI: 1.07, 1.58) and low muscle quality (HR 1.24, 95% CI: 1.03, 1.49) were associated with poor long-term survival, but not with the risk of recurrence and progression-free survival. Both low skeletal muscle index (SMI) (RR 1.49, 95% CI: 1.13, 1.98) and low muscle quality (RR 1.99, 95% CI: 1.04, 3.79) were associated with increased risk of complications. CONCLUSIONS: Both low skeletal muscle mass and low muscle quality showed significant association with poor long-term survival and an increased risk of complications. However, they do not have a significant association with the risk of recurrence and progression-free survival. There is a need for more prospective studies to confirm these associations.

[Changing trend of incidence and mortality of esophageal cancer during 2010-2016 in Henan Province, China].

Objective: To estimate the incidence and mortality of esophageal cancer in 2016 and their changing trend during 2010-2016 according to the cancer registration data in Henan province. Methods: The data quality including completeness, validity, and reliability of local registries which submitted the cancer registration data of 2016 were assessed according to the criteria of Guideline on Cancer Registration in China and IARC/IACR. Esophageal cancer cases (ICD10: C15) were extracted from the database, and the incidence and mortality stratified by gender, age, and areas (urban/rural) were calculated, the incidence and mortality of provincial cancer were estimated combined with provincial population data. China's 2000 census population and Segi's population were used to calculate the age-standardized rate. Joinpoint model was used to estimate the changing trend of age standardized incidence and mortality along with the calendar year. Results: Approximately 40.10 thousand new esophageal cancer cases were diagnosed in Henan in 2016, accounting for 13.46% of all new cancer cases, and it ranked the third among cancer of all sites. The crude incidence of esophageal cancer was 37.21/100 000 with an age-standardized incidence rate by China standard population (ASIRC) of 26.74/100 000 and an age-standardized incidence rate by world standard population (ASIRW) of 27.12/100 000. The incidence of esophageal cancer in males was higher than that in females, with the ASIRC of 34.53/100 000 and 19.19/100 000, respectively. It was higher in rural areas than that in urban areas, with the ASIRC of 28.13/100 000 and 20.90/100 000, respectively. About 29.30 thousand deaths of esophageal cancer in Henan in 2016, accounting for 15.61% of all cancer deaths in Henan, which ranked the third among cancer of all sites. The crude mortality rate was 27.14/100 000 with an age-standardized mortality rate by China standard population (ASMRC) of 18.74/100 000 and an age-standardized mortality rate by world standard population (ASMRW) of 18.78/100 000. The mortality in males was higher than that in females, with the ASMRC of 24.78/100 000 and 13.12/100 000, respectively. It was also higher in rural areas than that in urban areas, with the ASMRC of 19.48/100 000 and 15.73/100 000, respectively. The ASIRC and ASMRC were declining with annual percent change (APC) of 3.12% (APC=-3.12%; 95%CI: -5.30%, -0.90%; P=0.015) and 2.47% (APC=-2.47%; 95%CI: -4.70%, -0.20%; P=0.039) during 2010-2016. However, the significant declining trend was only observed in rural areas in Henan, and the changing trend was same between males and females. Conclusions: The incidence and mortality of esophageal cancer are declining since 2010, however, the disease burden remains large in Henan. Therefore, comprehensive prevention and control efforts should be strengthened according to its epidemic characteristics and risk factors.

[Changing trend of incidence and mortality of stomach cancer during 2010-2016 in Henan Province, China].

Objective: To estimate stomach cancer incidence and mortality in Henan, 2016 and analyze the trend of stomach cancer incidence and mortality from 2010 to 2016. Methods: Stomach cancer related data in 2016 was extracted from Henan cancer registration and follow-up system. All data were qualified in validity, reliability and completeness according to the Guideline on Cancer Registration in China and International Agency for Research on Cancer (IARC/IACR). The incidence and mortality of stomach cancer were estimated by areas, gender and age based on the quality data and the registered population data of Henan province in 2016. The epidemic trend of stomach cancer was also been evaluated based on the age-standardized incidence and mortality by Chinese population (ASR China) from 2010 to 2016. Results: In 2016, the estimated incident cases of stomach cancer were 44 311. The incidence was 41.07/100 000, ASR China was 30.17/100 000, ASR by world population (ASR world) was 30.36/100 000, and the cumulative incidence rate was 3.84%. The incidences of male and female were 55.65/100 000 and 25.35/100 000, respectively. Meanwhile, 32 927 people died of stomach cancer in Henan. The mortality was 30.52/100 000, ASR China was 21.45/100 000, ASR world was 21.54/100 000, and the cumulative mortality was 2.53%. From 2010 to 2016, both the ASR China for incidence and mortality of stomach cancer in Henan showed a steady downward trend. In rural, the ASR China for incidence and mortality decreased rapidly, while the stable trend was observed in urban. Nevertheless, the incidence and mortality of stomach cancer in rural were still higher than those in urban. Conclusions: The incidence and mortality of stomach cancer in Henan province showed steadily declining trend from 2010 to 2016, and the geographical distribution difference between rural and urban areas was gradually narrowing. However, the disease burden was still high in 2016.

[Acceptance evaluation of urine self-sampling, vaginal self-sampling and physician sampling in cervical cancer screening].

Objective: To compare the acceptance of urine self-sampling, vaginal self-sampling and physician sampling in cervical cancer screening. Methods: Questionnaire surveys were conducted in women attending cervical cancer screening in Xiangyuan County and Yangcheng County in Shanxi Province from July to November 2015 and from October to December 2018. Both surveys included the following three parts: feelings and acceptance of women in urine self-sampling, vaginal self-sampling and physician sampling, and the ease evaluation of two self-samplings. Results: There were 3 485 questionnaires were collected finally. The proportions of women felt embarrassed in urine self-sampling, vaginal self-sampling and physician sampling were 2.3% (81/3 472), 7.1% (247/3 472) and 8.6% (299/3 472), and 1.3% (46/3 472), 9.7%(3 37/3 472) and 14.9% (518/3 472) of women felt uncomfortable, and 1.0% (35/3 469), 8.5% (295/3 469) and 15.3% (531/3 469) felt pain, and 98.3% (3 334/3 393), 96.3% (3 267/3 393) and 99.0% (3 360/3 393) thought that the sampling were conducted properly (P<0.05). Based on the feeling during sample collection, 61.2% (1 876/3 064), 39.1% (1 199/3 064) and 66.5% (2 037/3 064) women were willing to use the corresponded sampling methods in cervical cancer screening, respectively (P<0.05). If the accuracy was the same, there were 31.8% (1 109/3 485) women preferred self-sampling and 68.2% (2 376/3 485) preferred physician sampling for cervical cancer screening. Meanwhile, 23.5% (820/3 482) preferred vaginal self-sampling in comparison with 76.5% (2 662/3 482) for urine self-sampling in cervical cancer screening. 86.5% (3 007/3 478) of the women thought urine self-sampling was very easy, comparing 40.9% (1 423/3 478) for vaginal self-sampling. Conclusions: The bad feeling of women during urine self-sampling is less common and less serious than those during physician sampling and vaginal self-sampling, and the acceptance for physician sampling is highest, following by urine self-sampling and vaginal self-sampling. Urine self-sampling is much easier than vaginal self-sampling.

[The characteristics of digital subtraction angiography and the analysis of the therapeutic effect of interventional therapy on hemorrhage after partial nephrectomy].

Objective: To investigate the digital subtraction angiography (DSA) manifestations of hemorrhage after partial nephrectomy (PN) for renal tumors, and the safety and efficacy of interventional therapy. Methods: The clinical data, DSA images and interventional treatment of 15 patients undergoing emergency interventional therapy due to hemorrhage after PN in the Department of Interventional Therapy of Beijing University Cancer Hospital from January 2015 to January 2019 were retrospectively analyzed. The DSA manifestations, safety and prognosis of interventional therapy were summarized. Results: Among the 15 patients, 10 had abdominal hemorrhage, 2 had abdominal hemorrhage with hematuria, and 3 had hematuria, all of which occurred 6 hours to 7 days after the operation. The DSA manifestations included contrast medium overflow, local artery wall unevenness, local artery branch expansion, pseudoaneurysm formation and renal arteriovenous fistula. There were 14 survivors, followed up for 6-12 months, and further no bleeding occurred. The success rate of interventional therapy was 100% (14/14), and the success rate of hemostasis was 100% (14/14). One patient died of asphyxia due to intraoperative nausea and vomiting. The creatinine levels were (76.0±11.0), (80.0±9.0), (76.0±9.0) µmol/L before operation, 1 week after operation and 1 month after operation respectively. There was no significant difference in creatinine levels among the three groups (F=3.171, P=0.058). The levels of urea nitrogen before operation, 1 week after operation and 1 month after operation were (6.80±1.04), (7.13±0.89), (6.81±0.79) µmol/L respectively. There was significant difference among the three groups (F=4.322, P=0.024). The level of urea nitrogen at 1 month after operation was significantly lower than that at 1 week after operation (P=0.016), but there was no difference compared with that before operation (P=1.000). Conclusion: Contrast medium overflow, pseudoaneurysm formation and renal arteriovenous fistula were the main manifestations of DSA hemorrhage after PN. Interventional therapy was safe and effective and might be a priority in cases of hemorrhage after PN.

[Analysis on influencing factors on HIV testing behaviors in some foreigners in Guangzhou].

Objective: To analyze the influencing factors of HIV testing behavior among foreigners in Guangzhou, and provide a reference for promoting HIV testing and intervention services in foreigners. Methods: A cross-sectional survey was conducted from November 2019 to January 2020 among 230 eligible foreigners selected in Guangzhou through convenient sampling method in the form of one-to-one questionnaire interview, HIV testing behaviors and related influencing factors were described and analyzed with χ(2) test and multivariate logistic regression model. Results: A total of 230 foreigners were included in the study, in whom 156 were males (67.8%), 217 were from Africa (94.3%), 138 (60.0%) were with education level of university and above, 169 (73.5%) were in business, 47 (20.4%) were students, 132 (57.4%) had cumulative residence >12 months in Guangzhou and 113 (49.1%) had health examination in China in the past 6 months. In these foreigners, the rate of AIDS-related knowledge awareness was 76.0%, the proportion of condom use in the last sex was 39.6%(89/225) and 47.8%(110/230) received HIV tests in the past 3 months. Multivariate logistic regression analysis showed, the influencing factors of HIV testing behavior in the past 3 months included cumulative residence ≤12 months (compared with cumulative residence >12 months, OR=3.12, 95%CI: 1.49-6.56), health examination in China in the past 6 months (compared to those who had not, OR=2.53, 95%CI: 1.17-5.48), awareness of HIV testing and consulting service in Guangzhou (compared with those who had no awareness, OR=3.65, 95%CI: 1.71-7.78). Conclusions: Some foreigners in Guangzhou had relatively low rates of HIV testing and condom use. Those who pay attention to their own health status and know well about HIV testing and consulting services are more likely to receive HIV testing, foreigners in Guangzhou should be encouraged to receive HIV testing. Their self-testing awareness should be improved.

[Performance of high-risk HPV typing test in early diagnosis of cervical cancer].

Objective: To evaluate the performance of High-risk HPV typing detection in cervical cancer screening. Methods: A total of 3 231 women were recruited as the subjects of cervical cancer screening from Jiyuan city of Henan provinces from June to July 2017. All women underwent HPV DNA test. The women with cytological examination ≥ASCUS or cytological examination negative and HPV 16/18 positive underwent colposcopy biopsy and pathological examination. Pathological diagnosis was used as the gold standard. Sensitivity, specificity, positive predictive value (PPV), negative predictive value(NPV) as well as corresponding 95% confidence interval (CI) of HR-HPV and HPV 16/18 were calculated. Results: The mean age of 3 231 subjects selected in this study was 46.84±10.00 (21-64) years old. 524 subjects had the positive results of HR-HPV, including 91 of HPV16 and 15 of HPV18. Pathological test result CIN2+ was the gold standard. The sensitivity and specificity of HR-HPV for cervical precancer lesions screening were 93.75 (95%CI: 79.85-98.27) and 84.56% (95%CI: 83.26-85.77), respectively. To compared with HR-HPV, HPV16/18 had low sensitivity (65.63%, 95%CI: 48.31-79.59)and higher specificity (97.44%, 95%CI: 96.83-97.93). After age stratification by age 30 and 45, the sensitivity of HPV 16/18 was same with HR-HPV (100%, 95%CI: 34.24-100.00), the specificity of HPV 16/18 was higher than HR-HPV (98.71%, 95%CI: 96.27-99.56 vs 84.48%, 95%CI: 79.27-88.58) in<30 age group.The sensitivity of HR-HPVin 30-45 and ≥45 age group were higher than HPV16/18 (85.71%, 95%CI: 48.69-97.43 vs 71.43%, 95%CI: 35.89-91.78, 95.65%, 95%CI: 79.01-99.23 vs 60.87%, 95%CI: 40.79-77.84), but the specificity werelower than HPV16/18 (86.89%, 95%CI: 84.58-88.90 vs 98.51%, 95%CI: 97.51-99.11、83.49%, 95%CI: 81.81-85.04 vs 96.80%, 95%CI: 95.94-97.48). Conclusions: HR-HPV detection has relatively high sensitivity and specificity in cervical cancer screening. For >30 years old women, HR-HPV is more recommended in cervical cancer screening. Therefore, HR-HPV detection is an effective method for cervical cancer screening.

A study of effect of lncRNA MVIH on sensitivity of gastric cancer cells to gemcitabine.

OBJECTIVE: To study the effect of expression of long non-coding ribonucleic acid (lncRNA) associated with microvascular invasion in hepatocellular carcinoma (MVIH) on the sensitivity of gastric cancer cells to gemcitabine. MATERIALS AND METHODS: BGC-823 cell lines were divided into control group (no treatment), low expression group (lentiviral transfection with sh-lncRNA MVIH), and high expression group (lentiviral transfection with lncRNA MVIH). The expression of lncRNA MVIH, the protein expressions of E-cadherin and Vimentin, and the differences in proliferation, migration, invasion, and apoptosis of gastric cancer cells were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western blotting, Cell counting kit-8 (CCK-8) assay, transwell assay, wound healing assay, methyl thiazolyl tetrazolium (MTT) assay, and flow cytometry. RESULTS: The results of RT-PCR revealed that compared with that in control group, the mRNA expression of lncRNA MVIH was significantly decreased in low expression group and markedly increased in high expression group. Also, there were statistically significant differences (p<0.05). The results of Western blotting showed that compared with those in control group and low expression group, the protein expression of E-cadherin was significantly decreased, while the protein expression of Vimentin was markedly increased in high expression group (p<0.05). The results of transwell assay manifested that the number of invading gastric cancer cells was the largest in high expression group at 48 h (p<0.05), significantly larger than that in control group and low expression group (p<0.05), while it was the smaller in low expression group. It was found through the wound healing assay that the migration ability of gastric cancer cells was enhanced in high expression group, markedly stronger than in control group and low expression group, while it significantly declined in low expression group compared to control group. Besides, the results of CCK-8 assay showed that compared with that in control group, the sensitivity of gastric cancer cells to gemcitabine was remarkably increased in low expression group (p<0.05), while it significantly declined in high expression group (p<0.05). Finally, according to the flow cytometry, the apoptosis rate of gastric cancer cells was markedly higher in low expression group than that in control group and high expression group (p<0.05), indicating that the low expression of lncRNA MVIH can promote the apoptosis of gastric cancer cells. CONCLUSIONS: Reducing the expression of lncRNA MVIH can significantly lower the migration and invasion ability of gastric cancer cells and raise the sensitivity of gastric cancer cells to gemcitabine.

Activation of liver X receptor promotes fatty acid synthesis in goat mammary epithelial cells via modulation of SREBP1 expression.

In bovine mammary tissue and cells, liver X receptor (LXR) regulates lipid synthesis mainly via transactivation of the transcription factor sterol regulatory element binding protein 1 (SREBP1). In the present work, we investigated the role of LXR in controlling lipid synthesis via transactivation of SREBP1 in goat primary mammary cells (GMEC). The GMEC were treated with a synthetic agonist of LXR, T0901317, and transactivation and transcription of SREBP1, expression of lipogenic genes, and fatty acid profiling and triacylglycerol (TAG) content of the cells were measured. A mild increase in the mRNA expression level of LXRα (NR1H3) was observed following treatment with different concentrations of T0901317, and a dose-dependent increase in mRNA and transactivation of SREBP1 was detected. Activation of LXR resulted in a significant increase in the mRNA expression of most of the measured genes related to de novo synthesis, desaturation, and transport of fatty acids; TAG synthesis; and transcription regulators. Compared with the control, total content of cellular TAG increased by more than 20% with T0901317 treatment. Furthermore, addition of T0901317 increased the proportion of unsaturated fatty acids (e.g., C16:1, C18:1, C20:1, and C22:1), and decreased the proportion of saturated fatty acids (e.g., C16:0, C18:0, C20:0, and C22:0). These results provide evidence that LXR regulates the expression and activity of SREBP1. Our results indicated that LXR participate in regulating the transcription of genes involved in milk fat synthesis in GMEC in an SREBP1-dependent fashion.

[The surgical treatment of ovarian cancer metastasis between liver and diaphragm: a report of 83 cases].

Objective: To explore the safety and feasibility of associating diaphragm resection and liver-diaphragmatic metastasis lesions resection for patients with advanced ovarian cancer. Methods: Retrospectively analysis 83 cases(98 times) of advanced ovarian cancer with liver-diaphragmatic metastasis between January 2012 and December 2016 at Department of Liver Surgery, Peking Union Medical College Hospital.The patients were aged from 19 to 75 years.Surgical procedure included metastatic lesions resection(43 times) and stripping(55 times). Operation status, post-operative complications, pathology results and follow-up of the patients were analyzed. Results: Fifteen patients received twice surgical treatment and 68 patients received one time surgical treatment. Postoperative hemorrhage in chest and between liver and diaphragm was not occurred in all cases.Dyspnea and low oxygen saturation were occurred in two cases of stripping patients and 1 case of metastatic lesions resection patients.Results of CT examination indicated that there was medium to large amount of ascites in right chests.The symptoms were relieved after placing thoracic closed drainage.Other patients were recovered smoothly.All patients were diagnosed as ovarian cancer by pathological examination. Conclusion: Associating diaphragm resection is safe and feasible for liver-diaphragmatic metastasis lesions from ovarian cancer.

[Predictors of rush popper use among 825 men who have sex with men in education in Guangdong Province].

Objective: To explore prevalence and predictors of the use of Rush poppers among men who have sex with men in education in Guangdong Province. Methods: An internet survey was conducted via a gay website from March to August 2014 and a sample of 833 male respondents over 16 years who had ever had anal sex with a male, studied in Guangdong Province and who had completed an online questionnaire was identified- of which 93.4% (n=825) provided valid responses and were included in the study. Respondents provided information on their socio-demographic characteristics, HIV testing history, sexual activity in last 3 months, Rush poppers use and time since first sexual encounter with a male. We used the Chi-squared test and multivariate logistic regression modeling to identify the predictors of Rush poppers use in this population group. Results: Among our sample of 825 MSM, whose ages ranged from 21-27 years, 14.8% (n=122) reported having used Rush poppers in the past 3 months. Rates of use were higher among those who had ever been tested for HIV (16.7% vs. 10.9%, χ2=4.80, P=0.028), had anal intercourse in the last 3 months (16.8% vs. 10.7%, χ2=5.48, P=0.019), had multiple sexual partners in the past 3 months (19.3% vs. 13.1%, χ2=5.14, P=0.023), had a casual partner (18.0% vs. 12.1%, χ2=5.68, P= 0.017) or had a sexually transmitted infection (23.4% vs. 14.0%, χ2=4.85, P=0.028). Those who reported having had their first sexual encounter with another male over 5 years prior had higher rates of use (18.0%) than for those who had been sexually active ≤1 year (10.7%) or 2-4 years (12.5%, χ2=6.41, P=0.041). HIV testing (OR: 1.70, 95% CI: 1.07-2.70), having a casual partner (OR: 1.67, 95% CI: 1.12-2.50) and being sexually active with other males for over 5 years (OR: 1.87, 95%CI: 1.05-3.31) were significantly associated with Rush poppers use. Conclusion: A large proportion of student MSM in Guangdong Province used Rush poppers. Proxy measures of sexual activity such as HIV testing, recent casual sexual encounters and duration of sexual activity were associated with their use.

[Application of " Internet Plus" AIDS prevention services among men who have sex with men in Guangzhou, China: results from 2010 to 2015].

Objective: To introduce the development strategy of " Internet Plus" AIDS prevention services and its implementation results from 2010 to 2015 in Guangzhou, China. Methods: A gay men's health column was created for an active website aimed at men who have sex with men(MSM), in collaboration with local community organizations and the Guangzhou CDC. We designed intervention tools(including scenario-based applications and HIV risk self-assessment systems)and an online HIV testing service platform, integrated with applied psychology and behavioral theory as well as the " Internet Plus" concept, to intervene in HIV infection risk factors among MSM. Data of clients who accessed the " Internet Plus" AIDS services from 2010 to 2015 were used to evaluate service operation. Six-year consecutive surveys, conducted between April and July of each service year, were collected using a national AIDS sentinel surveillance questionnaire. For each year of surveillance, information on HIV prevalence, HIV interventions received during the past year, unprotected anal intercourse in the past 6 months, and HIV testing in the past year were compared using the chi-squared(χ2)test, to roughly reflect the effect of"Internet Plus" AIDS prevention services. Results: As of 31 December 2015, a total of 34 395 MSM had received " Internet Plus" services and HIV testing. The number of MSM tested increased from 2 338 in 2010 to 8 054 in 2015. From 2010 to 2015, newly identified HIV cases in each year were 59, 166, 312, 283, 291, and 270, which accounted for 25.0%, 32.8%, 38.8%, 35.1%, 30.5%, and 23.2% of MSM HIV cases of Guangzhou, respectively. Sentinel surveillance data showed that during the study period, 3 047 MSM were investigated, with 405, 400, 401, 633, 608, and 600 each year, respectively. The proportion of participants who had received any HIV intervention during the past year was 74.3%(301), 70.8%(283), 83.3%(334), 85.0%(538), 69.1%(420), and 83.8%(503)each year, respectively(trend χ2=6.53, P=0.011). HIV testing done during the past year accounted for 44.0%(178), 44.3%(177), 49.4%(198), 53.4%(338), 56.1%(341), and 60.2%(361)each year, respectively(trend χ2=40.83, P<0.001). Unprotected anal intercourse in the past 6 months accounted for 59.3%(240), 62.0%(248), 56.6%(227), 57.0%(361), 48.4%(294), and 43.7%(262)each year, respectively(trend χ2=42.21, P<0.001). Conclusion: The"Internet Plus"AIDS prevention services in this study represent a manner to enhance traditional HIV prevention strategies. We found these services to be effective in implementation of the national AIDS control and prevention strategy, especially for the expansion of intervention, testing, and case identification among high-risk populations.

Liver X receptor α promotes the synthesis of monounsaturated fatty acids in goat mammary epithelial cells via the control of stearoyl-coenzyme A desaturase 1 in an SREBP-1-dependent manner.

Stearoyl-coenzyme A desaturase 1 (SCD1) is a pivotal enzyme in the biosynthesis of monounsaturated fatty acids (MUFA). It is tightly regulated by transcription factors that control lipogenesis. In nonruminants, liver X receptor α (LXRα) is a nuclear receptor and transcription factor that acts as a key sensor of cholesterol and lipid homeostasis. However, the mechanism whereby LXRα regulates the expression and transcriptional activity of SCD1 in ruminant mammary cells remains unknown. In this study with goat mammary epithelial cells (GMEC), the LXRα agonist T 4506585 (T09) markedly enhanced the mRNA expression of SCD1 and sterol regulatory element binding factor 1 (SREBF1). The concentrations of C16:1 and C18:1 and their desaturation indices also were increased by LXRα activation. However, knockdown of LXRα did not alter the mRNA expression of SCD1. Although SCD1 was repressed by SREBF1 knockdown, T09 significantly increased SCD1 expression. Further analysis revealed that the SCD1 promoter activity was activated by LXRα overexpression. The goat SCD1 promoter contains 2 LXR response elements (LXRE), 1 sterol response element (SRE), and 1 nuclear factor Y (NF-Y) binding site. Site-directed mutagenesis of LXRE1, LXRE2, or SRE alone did not eliminate the upregulation of SCD1 when LXRα was overexpressed. In contrast, when NF-Y alone or in combination with SRE was mutated simultaneously, the basal transcriptional activity of the SCD1 promoter was markedly decreased and did not respond to LXRα overexpression. Furthermore, when SREBF1 was knocked down, overexpression of LXRα did not affect the promoter activity of SCD1. Together, these data suggest that LXRα regulates the expression of SCD1 through increasing SREBP-1 abundance to promote interaction with SRE and NF-Y binding sites. The present study provides evidence that LXRα is involved in the synthesis of MUFA in the goat mammary gland through an indirect mechanism.

Short communication: Altered expression of specificity protein 1 impairs milk fat synthesis in goat mammary epithelial cells.

Specificity protein 1 (encoded by SP1) is a novel transcription factor important for the regulation of lipid metabolism and the normal function of various hormones in model organisms. Its potential role, if any, on ruminant milk fat is unknown. Despite the lower expression of the lipolysis-related gene ATGL (by 44 and 37% respectively), both the adenoviral overexpression and the silencing of SP1 [via short interfering (si)RNA] markedly reduced cellular triacylglycerol (TAG) content (by 28 and 25%, respectively), at least in part by decreasing the expression of DGAT1 (-36% in adenovirus treatment) and DGAT2 (-81 and -87%, respectively) that are involved in TAG synthesis. Consistent with the markedly lower expression of genes related to lipid droplet formation and secretion (TIP47 by 19 and 32%, and ADFP by 25 and 25%, respectively), cellular lipid droplet content was also decreased sharply, by 9 and 8.5%, respectively, after adenoviral overexpression of SP1 or its silencing via siRNA. Overall, the results underscored a potentially important role of SP1 in maintaining milk-fat droplet synthesis in goat mammary epithelial cells.

Thyroid hormone responsive (THRSP) promotes the synthesis of medium-chain fatty acids in goat mammary epithelial cells.

In nonruminants, thyroid hormone responsive (THRSP) is a crucial protein for cellular de novo lipogenesis. However, the role of THRSP in regulating the synthesis of milk fatty acid composition in goat mammary gland remains unknown. In the present study, we compared gene expression of THRSP among different goat tissues. Results revealed that THRSP had the highest expression in subcutaneous fat, and expression was higher during lactation compared with the dry period. Overexpression of THRSP upregulated the expression of fatty acid synthase (FASN), stearoyl-coenzyme A desaturase 1 (SCD1), diacylglycerol acyltransferase 2 (DGAT2), and glycerol-3-phosphate acyltransferase (GPAM) in goat mammary epithelial cells. In contrast, overexpression of THRSP led to downregulation of thrombospondin receptor (CD36) and had no effect on the expression of acetyl-coenzyme A carboxylase α (ACACA) and sterol regulatory element binding transcription factor1 (SREBF1). In addition, overexpressing THRSP in vitro resulted in a significant increase in triacylglycerol (TAG) concentration and the concentrations of C12:0 and C14:0. Taken together, these results highlight an important role of THRSP in regulating lipogenesis in goat mammary epithelial cells.

Overexpression of SREBP1 (sterol regulatory element binding protein 1) promotes de novo fatty acid synthesis and triacylglycerol accumulation in goat mammary epithelial cells.

Sterol regulatory element binding protein 1 (SREBP1; gene name SREBF1) is known to be the master regulator of lipid homeostasis in mammals, including milk fat synthesis. The major role of SREBP1 in controlling milk fat synthesis has been demonstrated in bovine mammary epithelial cells. Except for a demonstrated role in controlling the expression of FASN, a regulatory role of SREBP1 on milk fat synthesis is very likely, but has not yet been demonstrated in goat mammary epithelial cells (GMEC). To explore the regulatory function of SREBP1 on de novo fatty acids and triacylglycerol synthesis in GMEC, we overexpressed the mature form of SREBP1 (active NH2-terminal fragment) in GMEC using a recombinant adenovirus vector (Ad-nSREBP1), with Ad-GFP (recombinant adenovirus of green fluorescent protein) as control, and infected the GMEC for 48 h. In infected cells, we assessed the expression of 20 genes related to milk fat synthesis using real time-quantitative PCR, the protein abundance of SREBP1 and FASN by Western blot, the production of triacylglycerol, and the fatty acid profile. Expression of SREBF1 was modest in mammary compared with the other tissues in dairy goats but its expression increased approximately 30-fold from pregnancy to lactation. The overexpression of the mature form of SREBP1 was confirmed by >200-fold higher expression of SREBF1 in Ad-nSREBP1 compared with Ad-GFP. We observed no changes in amount of the precursor form of SREBP1 protein but a >10-fold increase of the mature form of SREBP1 protein with Ad-nSREBP1. Compared with Ad-GFP cells (control), Ad-nSREBP1 cells had a significant increase in expression of genes related to long-chain fatty acid activation (ACSL1), transport (FABP3), desaturation (SCD1), de novo synthesis of fatty acids (ACSS2, ACLY, IDH1, ACACA, FASN, and ELOVL6), and transcriptional factors (NR1H3 and PPARG). We observed a >10-fold increase in expression of INSIG1 but SCAP was downregulated by Ad-nSREBP1. Among genes related to milk fat synthesis and lipid droplet formation, only LPIN1 and DGAT1 were upregulated by Ad-nSREBP1. Compared with the Ad-GFP, the cellular triacylglycerol content was higher and the percentage of C16:0 and C18:1 increased, whereas that of C16:1, C18:0, and C18:2 decreased in Ad-nSREBP1 cells. Overall, the data provide strong support for a central role of SREBP1 in the regulation of milk fat synthesis in goat mammary cells.

Sterol regulatory element binding protein-1 (SREBP-1)c promoter: Characterization and transcriptional regulation by mature SREBP-1 and liver X receptor α in goat mammary epithelial cells.

Sterol regulatory element binding protein-1 (SREBP-1) is a key transcription factor that regulates lipogenesis in rodent liver. Two isoforms (SREBP-1a and SREBP-1c) of SREBP-1 are transcribed by an alternative promoter on the same gene (SREBF1), and the isoforms differ only in their first exon. Although the regulatory effects of SREBP-1 on lipid and milk fat synthesis have received much attention in ruminants, SREBP-1c promoter and its regulatory mechanisms have not been characterized in the goat. In the present study, we cloned and sequenced a 2,012-bp fragment of the SREBP-1c 5'-flanking region from goat genomic DNA. A luciferase reporter assay revealed that SREBP-1c is transcriptionally activated by the liver X receptor α (LXRα) agonist T0901317, and is decreased by SREBP-1 small interfering (si)RNA. A 5' deletion analysis revealed a core promoter region located -395 to +1 bp upstream of the transcriptional start site (TSS). Site-directed mutagenesis of LXRα binding elements (LXRE1 and LXRE2) and sterol regulatory elements (SRE1 and SRE2) revealed that the full effects of T 4506585 require the presence of both LXRE and SRE. We also characterized a new SRE (SRE1) and demonstrated a direct role of SREBP-1 (auto-loop regulation) in maintaining its basal transcription activity. Results suggest that goat SREBP-1c gene is transcriptionally regulated by mature SREBP-1 (auto-loop circuit regulation) and LXRα in goat mammary epithelial cells.

Prognostic role of microRNA-100 in patients with bladder cancer.

We investigated the clinical significance and prognostic value of microRNA-100 (miR-100) in bladder cancer. Quantitative real-time polymerase chain reaction was used to analyze the expression of miR-100 in 92 pairs of human bladder cancer and adjacent normal tissue samples. Overall survival (OS) curves were plotted using the Kaplan-Meier method and were evaluated for statistical significance using a log-rank test. The significance of different variables with respect to survival was analyzed using the multivariate Cox proportional hazard model. The miR-100 expression level was significantly lower in bladder cancer tissues than in normal adjacent tissues (mean ± SD: 1.49 ± 0.52 vs 2.79 ± 0.59, P < 0.05). A low miR-100 expression level was correlated with tumor stage (P = 0.023), tumor grade (P = 0.031), and regional lymph node involvement (P = 0.16). Kaplan-Meier analysis with log-rank test indicated that low miR-100 expression had a significant impact on OS (35.1 vs 75.3%; P = 0.004). Multivariate analysis revealed that the miR-100 expression level was an independent prognostic factor for OS (HR = 2.768, 95%CI = 1.287-8.992; P = 0.009) in bladder cancer patients. The present study demonstrated that the downregulation of miR-100 was associated with advanced clinical features and poor prognosis for bladder cancer patients, suggesting that miR-100 downregulation may be used as an unfavorable prognostic biomarker in bladder cancer.

MicroRNA-24 can control triacylglycerol synthesis in goat mammary epithelial cells by targeting the fatty acid synthase gene.

In nonruminants it has been demonstrated that microRNA-24 (miR-24) is involved in preadipocyte differentiation, hepatic lipid, and plasma triacylglycerol synthesis. However, its role in ruminant mammary gland remains unclear. In this study we measured miR-24 expression in goat mammary gland tissue at 4 different stages of lactation and observed that it had highest expression at peak lactation when compared with the dry period. Overexpression or downregulation of miR-24 in goat mammary epithelial cells (GMEC) strongly affected fatty acid profiles; in particular, miR-24 enhanced unsaturated fatty acid concentration. Additional effects of miR-24 included changes in triacylglycerol content and the expression of fatty acid synthase, sterol regulatory element binding transcription protein 1, stearoyl-CoA desaturase, glycerol-3-phosphate acyltransferase mitochondrial, and acetyl-CoA carboxylase. Luciferase reporter assay confirmed that fatty acid synthase is a target of miR-24. Taken together, these results not only highlight the physiological importance of miR-24 in fatty acid metabolism in GMEC, but also laid the foundation for further research on regulatory mechanisms among miR-24 and other microRNA expressed in GMEC.

Adipocyte differentiation-related protein promotes lipid accumulation in goat mammary epithelial cells.

Milk fat originates from the secretion of cytosolic lipid droplets (CLD) synthesized within mammary epithelial cells. Adipocyte differentiation-related protein (ADRP; gene symbol PLIN2) is a CLD-binding protein that is crucial for synthesis of mature CLD. Our hypothesis was that ADRP regulates CLD production and metabolism in goat mammary epithelial cells (GMEC) and thus plays a role in determining milk fat content. To understand the role of ADRP in ruminant milk fat metabolism, ADRP (PLIN2) was overexpressed or knocked down in GMEC using an adenovirus system. Immunocytochemical staining revealed that ADRP localized to the surface of CLD. Supplementation with oleic acid (OA) enhanced its colocalization with CLD surface and enhanced lipid accumulation. Overexpression of ADRP increased lipid accumulation and the concentration of triacylglycerol in GMEC. In contrast, morphological examination revealed that knockdown of ADRP decreased lipid accumulation even when OA was supplemented. This response was confirmed by the reduction in mass of cellular TG when ADRP was knocked down. The fact that knockdown of ADRP did not completely eliminate lipid accumulation at a morphological level in GMEC without OA suggests that some other compensatory factors may also aid in the process of CLD formation. The ADRP reversed the decrease of CLD accumulation induced by adipose triglyceride lipase. This is highly suggestive of ADRP promoting triacylglycerol stability within CLD by preventing access to adipose triglyceride lipase. Collectively, these data provide direct in vitro evidence that ADRP plays a key role in CLD formation and stability in GMEC.