Effectiveness and safety of REBACIN as a non-invasive intervention for persistent high-risk human papillomavirus infection: A real-world prospective multicenter cohort study.

BACKGROUND: We previously reported that REBACIN effectively eliminates persistent high-risk human papillomavirus (hrHPV) infection. Here, we conducted a prospective multicenter cohort study to evaluate the safety and effectiveness of REBACIN, taking into account factors such as specific hrHPV subtype and patient's age. METHODS: According to inclusion/exclusion criteria and participant willingness, 3252 patients were divided into REBACIN group while 249 patients into control group. Patients in REBACIN group received one course treatment of intravaginal administration of REBACIN while no treatment in control group. After drug withdrawal, participants in both groups were followed up. RESULTS: The clearance rate of persistent hrHPV infection in REBACIN group was 60.64%, compared to 20.08% in control group. Specifically, the clearance rates for single-type infection of HPV16 or HPV18 were 70.62% and 69.23%, respectively, which was higher than that of HPV52 (59.04%) or HPV58 (62.64%). In addition, the single, double, and triple/triple+ infections had a clearance rate of 65.70%, 53.31%, and 38.30%, respectively. Moreover, 1635 patients under 40 years old had a clearance rate of 65.14%, while it was 55.08% for 1447 patients over 40 years old. No serious adverse effects were found. CONCLUSION: This study confirmed that REBACIN can effectively and safely eliminate persistent hrHPV infection, which the clearance rate of HPV16/18 is higher than that of HPV52/58, the clearance rate of single-type infection is higher than that of multiple-type infections, and the clearance rate in young patients is higher than that in elder patients, providing a guidance for REBACIN application in clearing hrHPV persistent infection in real-world settings. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry Registration Number: ChiCTR1800015617 http://www.chictr.org.cn/showproj.aspx?proj=26529 Date of Registration: 2018-04-11.

Internet use time and mental health among rural adolescents in China: A longitudinal study.

BACKGROUND: The digital divide between urban and rural adolescents is widening. Many existing studies have found an association between internet use and adolescent mental health, but few use longitudinal data to focus on rural adolescents. We aimed to identify the causal relationships between internet use time and mental health in Chinese rural adolescents. METHODS: Using a sample of 3694 participants (aged 10-19) from the 2018-2020 China Family Panel Survey (CFPS). Fixed effects model, mediating effect model and instrumental variables method was used to evaluate the causal relationships between internet use time and mental health. RESULTS: We find that more time spent on the internet has a significant negative effect on participants' mental health. This negative impact is stronger in female and senior students groups. Mediating effects analysis suggests that more time spent on the internet increase risk of mental health problems by reducing sleep duration and parent-adolescent communication. Further analysis find that online learning and online shopping is associated with higher depression scores, while online entertainment with lower depression scores. LIMITATIONS: The data do not investigate the specific time spent on internet activities (e.g., learning, shopping, and entertainment), and the long-term impacts of internet use time and mental health have not been tested. CONCLUSIONS: Internet use time has a significant negative impact on mental health by crowding out sleep duration and parent-adolescent communication. The results provide an empirical reference for the prevention and intervention of mental disorders in adolescents.

Effect of the SIRT3-AMPK/PPAR pathway on invasion and migration of cervical cancer cells.

SIRT3 is a mitochondrial deacetylation protein that can promote the invasion and migration of cancer cells. We explored the effects of SIRT3 regulation of the AMPK/PPAR signaling pathway on triglycerides and the invasion and metastasis of cervical cancer cells. Immunohistochemical methods were used to detect SIRT3. The expression of AMPK and PPAR proteins in different cervical lesions was analyzed in combination with clinicopathological parameters. qRT-PCR and western blotting were used to determine the expression levels of SIRT3 in the C33a and SiHa cervical cancer cell lines. To observe the effects of altering SIRT3 levels by lentivirus transfection and the consequent changes in AMPK and PPAR protein expression, oil red O staining was used to determine intracellular triglycerides, and scratch assays and Transwell chamber experiments were performed to evaluate cervical cancer cell migration and invasion. Our data indicate that SIRT3, AMPK, and PPAR protein expression levels show an increasing trend with cervical lesion severity and are related to the degree of lymph node metastasis and differentiation; moreover, increased expression of SIRT3 can promote the expression of AMPK and PPAR proteins, is beneficial to the formation of intracellular neutral fat, and enhances the ability of cells to metastasize and invade. Our results suggest that SIRT3 activates AMPK/PPAR signaling pathways involved in cancer lipid metabolism and promotes metastasis and cell invasion.

SIRT3 promotes the invasion and metastasis of cervical cancer cells by regulating fatty acid synthase.

Sirtuin 3 (SIRT3) modulates mitochondria-localized processes and is implicated in the metabolic reprogramming of cancer cells, especially fatty acid (FA) synthesis. However, the relationship between SIRT3 and aberrant lipid synthesis in cervical cancer remains unclear. Here, we investigated the clinical relevance of SIRT3 expression in cervical squamous cell carcinoma (CSCC), cervical intraepithelial neoplasia (CIN), and normal tissues. To analyze the role of SIRT3 in CCSC in vitro, endogenous SIRT3 levels were up- and down-regulated in SiHa and C33a cells, respectively, via lentiviral-based transfection. Levels were quantified using qRT-PCR. Acetylation levels for acetyl-coA carboxylase (ACC1) were measured with the anti-acetyllysine antibody. Knockdown of SIRT3 reduced levels of cellular lipid content in cells. To investigate the role of SIRT3 in cell proliferation, nude mice were xenografted with SIRT3-overexpressing or SIRT3-knockdown CCSC cells. Overall, the results demonstrate that SIRT3 significantly contributed to the reprogramming of FA synthesis in CCSC by up-regulating ACC1 to promote de novo lipogenesis by SIRT3 deacetylation. Moreover, the findings show that the SIRT3-mediated regulation of FA synthesis played a critical role in the proliferation and metastasis of CCSC cells, suggesting that SIRT3 has therapeutic potential in CCSC treatment.

Over-expression of prolyl isomerase Pin1 promotes cervical tumorigenesis and metastasis.

Overexpression of the prolyl isomerase PIN1 is involved in tumorigenesis, but the role of PIN1 in cervical cancer is unclear. In this study, we examined PIN1 protein expression by immunohistochemistry in 221 paraffin-embedded samples from cervical cancer patients, cervical intraepithelial neoplasia patients, and control tissues, and found that high expression of PIN1 was significantly associated with lymph node metastasis (P=0.002), advanced stage according to the International Federation of Gynecology and Obstetrics guidelines (P=0.026). When endogenous PIN1 expression was knocked down using siRNA, cell proliferation, colony formation, migration, and invasion were inhibited in the SiHa cervical cancer cell line. Additionally, PIN1 knockdown increased E-cadherin and ß-catenin expression, and decreased expression of N-cadherin and Vimentin, suggesting that PIN1 can promote epithelial-mesenchymal transition (EMT). These results indicate that the Overexpression of the prolyl isomerase PIN1 in cervical cancer indicates tumor-Promotive properties of PIN1 that may be a marker of poor prognosis in cervical cancer patients, and the molecular determinants of epithelial polarity which have tumorigenesis enhancing impact, might through EMT.

Functional Role of NRF2 in Cervical Carcinogenesis.

Nuclear factor erythroid-2-related factor 2 (NFE2L2) is a transcription factor associated with resistance to chemotherapy and increased tumor growth. NRF2 is repressed by the inhibitor Keap1. The Keap1-NRF2 pathway is dysfunctional in multiple tumor types. Among Uighur women, the incidence of cervical squamous cell carcinoma (CSCC) and cervical intraepithelial neoplasia (CIN) was associated with elevated nuclear expression of NRF2 and decreased cytoplasmic expression of Keap1. Up-regulation of nuclear NRF2 was significantly associated with reduced cytoplasmic Keap1 expression. NRF2 positivity and Keap1 negativity were frequently associated with more advanced tumors (i.e., higher histological grade, lymph node involvement, and higher tumor stages) (p<0.05 for all). Methylated CpG islands in the Keap1 gene promoter in cervical cancer tissue were identified using MassARRAY. Moreover, promoter hypermethylation of this gene was significantly associated with decreased protein expression and increased nuclear NRF2 expression in cervical cancer tissues. Overexpression and knockdown of NRF2 in CSCC cell lines showed that NRF2 promotes proliferation, inhibits apoptosis, and enhances migration and invasion. These studies support the concept that epigenetic changes regulate expression of Keap1 in cervical cancer tissues. The association of NRF2 expression with aggressive tumor behavior suggests that NRF2 may be a marker of poor prognosis in patients with cervical cancer.

Epigenetic regulation of human riboflavin transporter 2(hRFT2) in cervical cancers from Uighur women.

In the present study, we studied the hypermethylation of the human riboflavin transporter 2 (hRFT2) gene and regulation of protein expression in biopsies from resected tissues from Uighur cervical squamous cell carcinoma (CSCC) patients and their neighboring normal tissues. hRFT2 gene promoter region methylation sequences were mapped in cervical cancer cell line SiHa by bisulfite-sequencing PCR and quantitative detection of methylated DNA from 30 pairs of Uighur's CSCCs and adjacent normal tissues by MassARRAY (Sequenom, San Diego, CA, USA) and hRFT2 protein expression was analyzed by immunohistochemistry. In SiHa, we identified 2 CG sites methylated from all of 12CpG sites of the hRFT2 gene. Analysis of the data from quantitative analysis of single CpG site methylation by Sequenom MassARRAY platform showed that the methylation level between two CpG sites (CpG 2 and CpG 3) from CpG 1~12 showed significant differences between CSCC and neighboring normal tissues. However, the methylation level of whole target CpG fragments demonstrated no significant variation between CSCC (0.476 ± 0.020) and neighboring normal tissues (0.401 ± 0.019, p>0.05). There was a tendency for translocation the hRFT2 proteins from cytoplasm/membrane to nucleus in CSCC with increase in methylation of CpG 2 and CpG 3 in hRFT2gene promoter regions, which may relate to the genesis of CSCC. Our results suggested that epigenetic modifications are responsible for aberrant expression of the hRFT2 gene, and may help to understand mechanisms of cervical carcinogenesis.

Post-transcriptional and epigenetic regulation of antigen processing machinery (APM) components and HLA-I in cervical cancers from Uighur women.

Normal function of human leukocyte antigen class I (HLA-I) and antigen processing machinery (APM) proteins is required for T cell-mediated anti-tumor or antiviral immunity, whereas the tumor survival indicates a failure of the host in immune surveillance associated with the dysfunction in antigen presentation, mainly due to the deregulation in HLA-I and APM expression or function. The posttranscriptional regulation of HLA-I and APM expression may associate with epigenetic modifications in cancer development which was not described so far. Here we showed that the development of cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) in Uighur women was accompanied with the partial or total loss of protein expression of HLA-I, ß2-m and APM components, including the transporter associated with antigen processing (TAP1/2), low molecular mass protein (LMP2, LMP7), endoplasmic reticulum aminopeptidase 1(ERAP1), chaperone molecules include calreticulin (CLR), calnexin (CNX) and ERp57, and this was proved again by analysis of transcription of the same genes in addition to three genes HLA-A, B and C coding for HLA-I. By bisulfite sequencing approach, we identified target CpG islands methylated at the gene promoter region of TAP1, TAP2, LMP7, tapasin and ERp57 in cervical carcinoma cells. Further analysis of CpG site specific methylation of these genes in cases of CSCC and CIN demonstrated an inverse correlation of altered CpG island methylation of TAP1, LMP7, and ERp57 with changes in protein expression. Moreover, promoter methylation of these genes was significantly higher in cases positive for human papillomavirus 16 (HPV 16) than negative ones. Our results suggested that epigenetic modifications are responsible for the aberrant expression of certain HLA-I and APM genes, and may help to understand unrevealed mechanisms of tumor escape from immune surveillance in cervical carcinogenesis.

Metabonomic signature analysis of cervical carcinoma and precancerous lesions in women by (1)H NMR spectroscopy.

(1)H nuclear magnetic resonance (NMR)-based metabonomics has been used to characterize the metabolic profiles of cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC). Principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were used to model the systematic variation related to patients with CIN or CSCC with healthy controls. Potential metabolic biomarkers were identified using database comparisons, and the one-way analysis of variance (ANOVA) test was used to examine the significance of the metabolites. Compared with plasma obtained from the healthy controls, plasma from patients with CIN had higher levels of very-low density lipoprotein (VLDL), acetone, unsaturated lipid and carnitine, together with lower levels of creatine, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine, glycine, acetylcysteine, myo-inositol, choline and glycoprotein. Plasma from patients with CSCC had higher levels of acetate and formate, together with lower levels of creatine, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine and tyrosine compared with the plasma of the healthy controls. In addition, compared with the plasma of patients with CIN, the plasma of CSCC patients had higher levels of acetate, formate, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine, tyrosine, acetylcysteine, myo-inositol, glycoprotein, α-glucose and ß-glucose, together with lower levels of acetone, unsaturated lipid and carnitine. Moreover, the profiles showed high feasibility and specificity by statistical analysis with OPLS-DA compared to the Thinprep cytology test (TCT) by setting the histopathological outcome as standard. The metabolic profile obtained for cervical cancer is significant, even for the precancerous disease. This suggests a systemic metabolic response to cancer, which may be used to identify potential early diagnostic biomarkers of the cancer and to establish clinical diagnostic methods.

[Relationship between TAP gene promoter methylation and cervical lesions with HPV infection in Uyghur women].

OBJECTIVE: To study the relationship between TAP (transporter associated with antigen processing) gene promoter regional methylation level and cervical lesions with HPV infection in Uyghur women. METHODS: A specialized software was used to design specific primers of CpG island fragments of TAP1 and TAP2 gene promoter for PCR amplification, bisulfitemodified SiHa cancer cell DNA for PCR amplification, cloning and sequencing analysis to obtain the relevant information on the gene base sequence methylation of CpG sites. Seventy-eight fresh cervical tissue samples from Uyghur women with cervicitis (number = 15), cervical intraepithelial neoplasia (CIN, number = 30) and cervical squamous cell carcinoma (number = 33) were collected. The methylation level of TAP1 and TAP2 gene promoter regions was detected using MassArray DNA technology. HPV infection status was determined by HPV gene chips. The relationship between CpG-island methylation of gene promoter regions and HPV infection was then analyzed. RESULTS: Each TAP1 and TAP2 gene corresponding target fragment contained 23 and 8 CpG sites. There were 5 and 8 CpG sites methylation occurred in SiHa cervical cancer cells genomic DNA respectively. The TAP1 methylation level increased steadily with the severity of cervical lesions. The methylation levels in cervical squamous cell carcinoma and CIN (0.048 ± 0.039 and 0.037 ± 0.026, respectively) were higher than that of normal cervical tissue (0.035 ± 0.029, P < 0.05). Although TAP2 gene methylation level also demonstrated similar changes, the difference however was not statistically significant (P > 0.05). HPV gene chip detected 13 HPV genotypes, with HPV16 infection rate being 66.7% (52/78). The methylated proportion of TAP1 positively correlated with HPV16 infection (χ(2) = 6.08, P = 0.039). CONCLUSION: TAP1 methylation is a remarkable phenomenon occurring in a range of cervical lesions and significantly associated with cervical HPV infection.

[Differential expressed genes in eutopic endometrium of endometriosis patients: comparison between Uygur and Han women in Xianjiang].

OBJECTIVE: To investigate the reason why the incidence of endometriosis (EM) is lower in the Uygur women than in the Han women. METHODS: Eutopic and ectopic endometrium samples were obtained by operation and biopsy from 26 EM patients, 10 Uygur women and 16 Han women and analyzed with a gene expression microarray containing the cDNAs of 22 000 human genes. Twenty-two women, 10 Uygur and 12 Han, were used as controls. RESULTS: Eleven differentially expressed genes, 7 up-regulated and 6 down-regulated, were screened out from the eutopic endometrium of the Uygur women with EM. 58 differential expressed genes were screened out from the in eutopic endometrium of the Han women with EM, 53 being up-regulated and 5 down-regulated. Five genes were screened out in both groups, 3 being up-regulated and 2 down-regulated. CONCLUSION: The number of differentially expressed genes of the Uygur women with EM is lower than that of the Han women with EM, which may be the cause of relative low incidence of EM among Uygur women.

[Clinical study of different forms of hysteroscopic surgery for endometrial polyps].

OBJECTIVE: To investigate the curative effects of different types of hysteroscopic surgery for endometrial polyps. METHODS: A total of 327 cases by different ways of hysteroscopic surgery for endometrial polyps from Nov 1999 to Nov 2004 were followed up. The mean age was (40 +/- 6) years. The mean follow-up was (3.0 +/- 0.6) years. Among 228 polyps patients in sexual maturity without desire of maintaining fertility, 53 (group A) underwent polypectomy with electrosurgical vaporization, and 175 cases (group B) did polypectomy with endometrial resection. Fifty-four (group C) cases (19 cases of infertility), who desired future childbearing, did polypectomy with endometrial resection of superficial layer near the polyps. Forty-five postmenopausal patients (group D) did polypectomy with endometrial coagulation. RESULTS: The time of operation: group A (15.1 +/- 0.8) second, group B (19.7 +/- 0.7) second, group C (20.9 +/- 0.7) second, and group D (22.1 +/- 0.8) second. None of the polyps recurred for the patients of groups A and D after operation, and the recurrent rate of groups B and C was 1.7% and 7.4%. There were no cases with amenorrhea in group C, who hoped to keep the function of fertility, but the recurrent rate of polyps was higher than other three groups. Of 19 cases of infertility, 14 cases became pregnant after the surgery. CONCLUSION: It is feasible to select different hysteroscopic surgery for endometrial polyps, according to different ages and the desire of childbearing of the patients.