Baicalin Antagonizes Prostate Cancer Stemness via Inhibiting Notch1/NF-κB Signaling Pathway.

OBJECTIVE: To investigate the molecular mechanisms underlying the effect of baicalin on prostate cancer (PCa) progression both in vivo and in vitro. METHODS: The in situ PCa stem cells (PCSCs)-injected xenograft tumor models were established in BALB/c nude mice. Tumor volume and weight were respectively checked after baicalin (100 mg/kg) treatment. Hematoxylin-eosin (HE) staining was used to observe the growth arrest and cell necrosis. mRNA expression levels of acetaldehyde dehydrogenase 1 (ALDH1), CD44, CD133 and Notch1 were determined by reverse transcription-polymerase chain reaction. Protein expression levels of ALDH1, CD44, CD133, Notch1, nuclear factor κB (NF-κB) P65 and NF-κB p-P65 were detected by Western blot. Expression and subcellular location of ALDH1, CD44, CD133, Notch1 and NF-κB p65 were detected by immunofluorescence analysis. In vitro, cell cycle distribution and cell apoptosis of PC3 PCSCs was assessed by flow cytometry after baicalin (125 µmol/L) treatment. The migration and invasion abilities of PCSCs were assessed using Transwell assays. Transmission electron microscopy scanning was utilized to observe the structure and autophagosome formation of baicalin-treated PCSCs. In addition, PCSCs were infected with lentiviruses expressing human Notch1. RESULTS: Compared with the control group, the tumor volume and weight were notably reduced in mice treated with 100 mg/kg baicalin (P<0.05 or P<0.01). Histopathological analysis showed that baicalin treatment significantly inhibited cell proliferation and promoted cell apoptosis. Furthermore, baicalin treatment reduced mRNA and protein expression levels of CD44, CD133, ALDH1, and Notch1 as well as the protein expression of NF-κB p-P65 in the xenograft tumor (P<0.01). In vitro, the cell proliferation of PCSCs was significantly attenuated after treatment with 125 µmol/L baicalin for 72 h (P<0.01). The cell migration and invasion rates were decreased following treatment with baicalin for 48 and 72 h (P<0.01). Baicalin notably induced cell apoptosis and seriously damaged the structure of PCSCs. The mRNA and protein expressions of CD133, CD44, ALDH1 and Notch1 in PCSCs were significantly downregulated following baicalin treatment (P<0.01). Importantly, the inhibitory effects of baicalin on PCa progression and stemness were reversed by Notch1 overexpression (P<0.05 or P<0.01). CONCLUSION: Mechanistically, baicalin exhibited a potential therapeutic effect on PCa via inhibiting the Notch1/NF-κB signaling pathway and its mediated cancer stemness.

Analgesic effect of electroacupuncture on bone cancer pain in rat model: the role of peripheral P2X3 receptor.

Upregulation of P2X3 receptor (P2X3R) has been strongly implicated in nociceptive signaling including bone cancer pain (BCP). The present study, using rat bone cancer model, aimed to explore the role of P2X3R in regulating rat pain behavior under the intervention of electroacupuncture (EA). The BCP model was successfully established by injection with MRMT-1 breast cancer cell into the medullary cavity of left tibia for 3 × 104 cells/3 µL PBS in rats as revealed by obvious bone destruction, decreased paw withdrawal thresholds (PWTs), and reduced paw withdrawal latencies (PWLs). Western blot analyses showed that P2X3R expression was significantly upregulated in ipsilateral lumbar 4-6 (L4-6) dorsal root ganglia (DRG), but the difference not seen in spinal cord dorsal horn (SCDH). With the in-depth study of P2X3R activation, we observed that intrathecal injection of P2X3R agonist α,ß-meATP aggravated MRMT-1 induced BCP, while injection of P2X3R inhibitor A-317491 alleviated pain. Subsequently, we demonstrated that BCP induced mechanical allodynia and thermal hyperalgesia were attenuated after EA treatment. Under EA treatment, total P2X3R protein expression in ipsilateral DRGs was decreased, and it is worth mentioning that decreased expression of P2X3R membrane protein, which indicated that both the expression and membrane trafficking of P2X3R were inhibited by EA. The immunofluorescence assay showed that EA stimulation exerted functions by reducing the expression of P2X3R-positive cells in ipsilateral DRGs of BCP rats. Ca2+ imaging analysis revealed that the EA stimulation decreased the percentage of α,ß-meATP responsive neurons in DRGs and inhibited calcium influx. Notably, the inhibitory effect of EA on mechanical allodynia and nociceptive flinches was abolished by intrathecal injection of α,ß-meATP. These findings demonstrated EA stimulation ameliorated mechanical allodynia and thermal hyperalgesia in rat model of MRMT-1-induced BCP. EA exerts analgesic effect on BCP by reducing the overexpression and functional activity of P2X3R in ipsilateral DRGs of BCP rats. Our work first demonstrates the critical and overall role of P2X3R in EA's analgesia against peripheral sensitization of MRMT-1-induced BCP and further supports EA as a potential therapeutic option for cancer pain in clinic.

A Population-based Study of Invasive Cervical Cancer Patients in Beijing: 1993-2008.

BACKGROUND: In recent years, the incidence of cervical cancer has been rising, particularly in young adults, as the second most common gynecological cancer in China. The aim of this study was to explore the incidence change and the epidemiological characteristics of cervical cancer in Beijing over the past 16 years. METHODS: All the cases of the study were limited to Beijing residents diagnosed with cervical cancer and registered in Beijing from January 1, 1993, to December 31, 2008. A total of 4100 patients with cervical cancer were obtained from the Statistics Database of Beijing Cancer Registry (BJCaR). According to the registered data, we retrospectively reviewed all original cases which we can acquired in reported hospital. Cervical situ cancer, cervical metastatic cancer, non-Beijing residents and repeatedly registered cases were excluded. Totally, 3641 registered cases were verified correctly. Meanwhile, we also collected the following data: Age, occupation, detected methods, histological type, and staging. The trends of incidence and mortality were analyzed by Joinpoint Regression Program 4.1.1.1 produced by National Cancer Institute (NCI, USA). The annual percent change (APC) was calculated using the Joinpoint regression model. RESULTS: The crude rates of incidence and mortality were 10.4 and 1.0 per 100,000 women, respectively during 1993 to 2008. The average WHO age-standardized incidence rates were 11.5 per 100,000 women. There was a decrease in incidence annually by 8.0% (P = 0.3) during 1993-1996 and a rapid increase annually by 18.9% after 1999 (P < 0.01). The median age was 67 years in 1993, but the median age decreased to 45 years in 2008. The peak of the age-specific incidence curve was at 40 years in the most recent period (2005-2008), which was 25-30 years earlier than that in previous periods (1993-1996). In the 2224 cases, the numbers of patients with stage I, II, III and IV were 910 (40.9%), 601 (27%), 542 (24.4%), 171 (7.7%), respectively. The percentage of patients with stage I was 7.6% (13/171) in 1993-1996, but the percentage increased to 51.6% (643/1247) in 2005-2008 (P < 0.01). Otherwise the percentage of advanced stage (stage III-IV) during the same period was dropped down significantly from 52.0% (89/171) to 22.5% (280/1247) (P < 0.01). Unemployed and housewife ranked first accounting for 27.3% of the total (607/2224). Urban low-income people such as worker ranked the second accounting for 17.0% (377/2224), the third place was farmer accounting for 14.0% (312/2224). Only 381 (17.1%, 381/2224) women in 2224 were first detected cervical cancer by routine screenings. Company staff (36.5%, 139/381), professional and technical personnel (22.6%, 86/381), national official (22.0%, 84/381) occupied the top three (total 81.1%) in the 381 patients detected cervical cancer by screening. CONCLUSIONS: The cervical cancer incidence has showed a continuous rise in Beijing since 1999. Government-led mass screening should target the low socioeconomic population primarily. Meanwhile the government should enhance public health education of cancer screening to increase the rate of screening.

[Study on subtype of human papillomavirus infection among aged 25 - 54 reproductive women in Beijing from 2006 to 2008].

OBJECTIVE: To investigate subtype of HPV infection among women at age of 25 to 54 years in Beijing. METHODS: From Sept. 2006 to Dec. 2008, 5552 reproductive women at age of 25 to 54 years classified into each 5-year as group were screened. Each participant completed a questionnaire and a sample of exfoliated cervical cells for liquid-based cytology and HPV subtype gene testing was performed. RESULTS: The overall prevalence of HPV was 6.68% (371/5552). High-risk HPV and Low-risk HPV prevalence were 5.76% (320/5552) and 2.00% (111/5552), respectively. The most common HPV types were HPV16 (2.61%, 145/5552), HPV58 (0.97%, 54/5552), HPV33 (0.85%, 47/5552), HPV43 (0.74%, 41/5552) and HPV56 (0.70%, 39/5552). There were two peaks of HPV prevalence: groups of 30 - 34 years and 40 - 44 years. CONCLUSION: The most common HPV subtypes in Beijing were HPV 16, 58, 33 and 43 and HPV prevalence showed a bi-modal age-specific curve.

[Present epidemiological status and clinical characteristics of cervical cancer in Beijing: a report of 1399 patients].

OBJECTIVE: To analyze the prevalence and clinical characteristics of hospitalized patients with cervical cancer over the last two decades in Beijing so as to provide scientific rationales for the management of cervical cancer. METHODS: Stratified cluster sampling was employed to analyze a total of 1399 invasive cervical cancer cases from different class hospitals in Beijing during the period of 1990 - 2009. RESULTS: (1) The number of cervical cancer cases had been rising over the last 2 decades. (2) The age of onset ranged from 17 to 88 years old; the number of patients with stages I, II, III and IV was 772 (57.1%), 380 (28.1%), 182 (13.5%) and 18 (1.3%) respectively; 1135 (83.1%) patients were of squamous cell carcinoma while 182 (13.3%) adenocarcinoma. The last two decades was divided into 4 groups of 5 years each. The average age gradually decreased from 58.2 to 46.0 years old (P < 0.01). The proportion of patients with stages I and II also increased from 69.6% (16/23) to 89.4% (530/856); There was no significant change in histopathologic type of cervical cancer. And squamous cell carcinoma of cervix remained the predominant type (P > 0.05). (3) About 26.0% (364/1399) of cases received no vagino-recto-abdominal examination while 3.4% (48/1399) of cases were not staged. CONCLUSION: Over the last two decades, the number of newly diagnosed cervical cancer cases has showed a rising trend in Beijing. And the proportions of young patients and patients with early disease have both increased. Some irregularities still exist in the diagnosis of cervical cancer. So we should pay more attention to the screening program and standardize the diagnosis, therapy and follow-up of cervical cancer.

[Analysis of high risk factors associated with cervical intraepithelial neoplasia in married women aged 25 - 54 years in Beijing between 2007 - 2008].

OBJECTIVE: to investigate high risk factors associated with cervical intraepithelial neoplasia (CIN) in married women aged 25 to 54 years in Beijing. METHODS: from Mar. 2007 to Sep. 2008, 6339 married women at age of 25 to 54 years were selected randomly by cross sectional survey in a total of 137 communities of 12 districts or counties in Beijing. The interview was carried out with unified questionnaires, gynecological examination. The cervical smear cytology and high-risk human papillomavirus (HR-HPV) infection of cervical secretion were detected. Women with abnormal cervical cytology underwent colposcopy and cervical biopsy. Odd ratio (OR) and 95% confidence interval (CI) of related high risk factors with CIN were studied by logistic regression analysis. RESULTS: among 6339 women, the prevalence rate of CIN including 4 squamous cell carcinoma (SCC) was 5.90% (374/6339). By multinomial regression analysis, HR-HPV infection (95%CI: 9.953 - 15.811), History of trichomonas vaginitis (95%CI: 1.046 - 2.104), oral contraceptives (95%CI: 1.087 - 1.806), age less than 45 years old (95%CI: 1.069 - 1.828) were related with CIN. CONCLUSION: infection rate of HR-HPV is an independent risk factor of CIN, however, the history of trichomonas vaginitis, oral contraceptives and age less than 45 years old are related risk factors of CIN.

[Epidemiological study on the prevalence of cervical intraepithelial neoplasia among married women aged 25-54 years in Beijing between 2007-2008].

OBJECTIVE: To investigate prevalence of cervical intraepithelial neoplasia (CIN) among the married women aged 25-54 years in Beijing. METHODS: With method of cross sectional survey, 6339 married women at age of 25 to 54 years were selected randomly in 137 communities of 12 districts or counties in Beijing from March 2007 to September 2008. The cervical smear cytology and high-risk human papillomavirus (HPV) infection of cervical secretion were detected. Women with abnormal cervical cytology were underwent colposcopy and cervical biopsy. RESULTS: Among 6339 women, 9.58% (607/6339) cases had abnormal cytological results, the colposcopy and cervical biopsy showed the rate of CIN was 5.84% (370/6339) in total selected women and 60.96% (370/607) in women with abnormal cervical cytology, including 4.65%(295/6339) in CIN I, 0.80% (51/6339) in CIN II, 0.38% (24/6339) in CIN III; 0.06% (4/6339) in early invasive carcinoma (SCC). Based on geographical distribution, the rate of cervical lesions was 4.46% (42/942) in urban areas, 6.27% (188/3000) in suburbs and 6.01% (144/2397) in outer suburbs (P > 0.05). CONCLUSION: It was found that the incidence of CIN was 5.84% in married women aged 25-54 years in Beijing, which did not show significant prevalence in urban, suburb and outer suburbs region.

[Analysis of correlation factors for cervical intraepithelial neoplasia in married women in Beijing, China].

OBJECTIVE: To investigate the characteristic of precancerous conditions of cervical intraepithelial neoplasia (CIN) and its related high-risk factors among the married women in Beijing, China. METHODS: Based upon the method of cross-sectional survey, 6339 married women at reproductive age from 25 to 54 years old were selected randomly in a total of 137 communities of 12 districts or counties in Beijing from March 2007 to September 2008. The interview was carried out with unified questionnaires and gynecological examinations. And the cervical Pap smear was used for liquid-based cytological examination and the cervical secretion for detection of high-risk human papillomavirus (HPV) DNA. Furthermore, the women with abnormal cervical cytology received colposcopy and cervical biopsy. RESULTS: (1) Among 6, 339 study subjects, 374 cases had CIN and the prevalence rate was 5.9%; (2) For the cases with cervical cytology positive results, cervical biopsy showed an elevated level of abnormal cytology and an increased incidence of cervical lesions; (3) The peak age of CIN prevalence was 30 to 34 years old and there was a high grade of cervical neoplasia; (4) Among the population, the infection rate of high-risk HPV was 9.9% and the infection rate of high-risk HPV in positive cytological group (41.2%) was significantly higher than that in negative cytological group (6.6%); (5) The infection rate and DNA load of high-risk HPV increased following the severity grade of CIN. CONCLUSION: In Beijing, married women at 30 to 34 years old are the high-risk group in CIN incidence and the infection of high-risk HPV is an independent risk factor. Liquid-base cytology combined with high-risk HPV DNA test is a viable method to discover CIN in time and prevent the incidence of cervical cancer.

[Study on the relations between high risk human papillomavirus and cervical intraepithelial neoplasia in married women from Beijing].

OBJECTIVE: To investigate the prevalence and relationship between high risk human papillomavirus (HR-HPV) and cervical intraepithelial neoplasia (CIN) in married women from Beijing. METHODS: From March 2007 to September 2008, a total of 6185 married women were sampled, covering 137 communities in 12 districts. The samples were screened by high-risk HPV DNA test (HC2) and cytological test. For those participants with cytological test results ≥ ASCUS, pathological tests were performed. An interview was also carried out with the same questionnaire. Results from the tests were inputted into the database twice using EpiData 3.0, reviewed, analyzed, using SPSS 15.0. RESULTS: (1) The prevalence rates of HR-HPV and CIN were 9.9% and 6.0%, respectively for the age group 25 to 54. (2) The peak age groups for HR-HPV and CIN prevalence rates were 30 to 34 years old. (3) The prevalence rates of positive cytology (40.3%) and CIN (30.4%) in HR-HPV positive female population were significantly higher than that in HR-HPV negative group. (4) Data from unconditional logistic regression analysis showed that, when comparing with the normal subjects, the risk odds ratios of HR-HPV with low grade CIN and cervical cancer/high grade CIN were 8.385 and 97.416 and the attributable risk proportions with these groups were 88.1% and 99.0%, respectively. CONCLUSION: HR-HPV infection seemed to be the main risk factor for CIN. Married women, from age group 30 - 34, were under the high risk group in both HR-HPV infection and CIN incidence.

[Prevalence and determinants of lower reproductive tract infections among women aged 25 - 54 years in Beijing].

OBJECTIVE: To determine the prevalence and determinants of the most commonly seen lower reproductive tract infections among women aged 25 - 54 years in Beijing. METHODS: The study population consisted of 6339 women aged 25 - 54 years in 137 communities of Beijing. Focus of this study was to understand the prevalence of the following diseases as:bacterial vaginosis, trichomoniasis and vulvovaginal candidiasis. In addition to their prevalence rates, a generalized equation for estimation was used to analyze those infection-associated factors. RESULTS: The overall infection prevalence in the lower reproductive tract was 11.4%, including bacterial vaginitis as 8.7%, trichomonads as 1.0% and vulvovaginal candidiasis as 1.7%. Factors which were found to be significantly associated with lower reproductive tract infections in women were age, profession, family income, number of sex partners and frequency of condom use during sexual contacts. In patients with bacterial vaginitis, both prevalence rates of cervical intraepithelial neoplasia and infection of human papillomavirus were high. CONCLUSION: The prevalence of the most commonly seen lower reproductive tract infections among women aged 25 - 54 years in Beijing was lower than other areas in China. Lower reproductive tract infections seemed to be related to 30 - 49 years of age, nongovernmental employee, poverty, higher number of sex partners and not using condoms during sexual contacts.

[Epidemiological study of high risk human papillomavirus infection in 25 to 54 years old married women in Beijing.].

OBJECTIVE: To investigate high risk human papillomavirus (HR-HPV) prevalence among married women in Beijing and to study the high risk factors. METHODS: During March 2007 to September 2008, a total of 6185 married women sampled from 137 communities in 12 districts were screened by HR-HPV DNA test and cytological test. The interview was carried out with unified questionnaires. The database was set up and twice entered in EpiData 3.0. After checked up, the data were analyzed in SPSS 15.0. RESULTS: (1) The HR-HPV infection rate was 9.89%. The HR-HPV infection rate of the city zone, the suburb and the exurb were 9.34%, 10.51% and 9.51% (P > 0.05). The HR-HPV infection rate of the native and the outlander were 9.53%, 11.30% (P < 0.05). (2) The age distribution of HR-HPV infection was that the rate was around 10% among 25 to 44 age groups, which was the highest (11.21%) in 30 to 34 age group; then the rate was descended as the age raising, the rate of 50 to 54 age group was the lowest (7.78%). (3) Multiple logistic regression showed that the related risk factors of HR-HPV infection mainly included 1000 RMB and above of family income per person per month, possessing more than 1 sexual partner of her husband, outlander and high levels of education. (4) The prevalence of cervical intraepithelial neoplasia (CIN) in HR-HPV positive group was significantly higher than that in HR-HPV negative group (29.76% vs 3.32%, P < 0.01). CONCLUSIONS: (1) The HR-HPV infection rate among aged 25 to 54 years was 9.9% and there was no significant difference in area distribution. (2) The high risk population which should strengthen screening was the married bearing-age women with high level of family income, outlander, high levels of education and her husband possessing more than 1 sexual partner. (3) HR-HPV infection is the main risk factor for CIN and cervical cancer, while does not provide a causal relationship with them. The high risk population should be checked regularly to understand the development of HR-HPV infection and CIN incidence.