CCT6A may act as a potential biomarker reflecting tumor size, lymphatic metastasis, FIGO stage, and prognosis in cervical cancer patients.

OBJECTIVE: Chaperonin-containing tailless complex polypeptide subunit 6A (CCT6A) is a critical regulator and newly identified clinical biomarker of several cancers, while its correlation with the clinical characteristics and prognosis of cervical cancer patients is unclear. Therefore, this study aimed to explore this issue. METHODS: Chaperonin-containing tailless complex polypeptide subunit 6A expression in tumor and tumor-adjacent tissues from 198 cervical cancer patients who underwent resection were detected by immunohistochemistry assay and reverse transcription-quantitative polymerase chain reaction. Besides, the clinicopathological features and survival data of cervical cancer patients were collected. RESULTS: Chaperonin-containing tailless complex polypeptide subunit 6A protein and mRNA levels were both increased in tumor tissues compared with tumor-adjacent tissues (both p < 0.001). Receiver operating characteristic curves showed that CCT6A protein (AUC: 0.774, 95% CI: 0.729-0.819) and mRNA levels (AUC: 0.904, 95% CI: 0.874-0.934) well discriminated tumor tissues from tumor-adjacent tissues. Besides, correlation analyses found that CCT6A protein and mRNA levels were positively correlated with lymph node metastasis and FIGO stage (all p < 0.05), apart from which CCT6A mRNA level was also positively associated with tumor size (p = 0.032). In addition, CCT6A protein and mRNA levels were negatively correlated with accumulating disease-free survival (both p < 0.05); meanwhile CCT6A mRNA level was negatively associated with accumulating overall survival as well (p = 0.010). CONCLUSION: Chaperonin-containing tailless complex polypeptide subunit 6A is elevated in tumor tissues, and its high expression associates with larger tumor size, lymph node metastasis, higher FIGO stage, and worse prognosis in cervical cancer patients.

MicroRNA­143­3p levels are reduced in the peripheral blood of patients with gestational diabetes mellitus and influences pancreatic ß­cell function and viability.

Pancreatic ß-cell dysfunction has been demonstrated to mediate key roles in the pathogenesis of gestational diabetes mellitus (GDM). Accumulating evidence has supported the functional involvement of microRNAs (miRNAs) in various types of diabetes, including GDM. However, the detailed biological effect of miRNAs in pancreatic ß-cell dysfunction remains poorly understood. In the present study, microarray data of miRNAs in the blood plasma of patients with GDM were retrieved from the Gene Expression Omnibus dataset under the accession number GSE98043. Reverse transcription-quantitative PCR (RT-qPCR) was performed to measure the expression levels of miR-143-3p in the blood plasma isolated from 30 female patients with GDM women and 30 healthy female individuals. Subsequently, murine pancreatic ß-cell line, MIN6 cells were treated with high glucose (HG) to construct in vitro cell models of GDM. miR-143-3p in HG-treated MIN6 cells was overexpressed or knocked down using miR-143-3p mimics and miR-143-3p inhibitor. Cell viability, insulin secretion and proinflammatory cytokine production were examined using CCK-8 and ELISA, respectively Cell apoptosis was measured by flow cytometry assay. The protein expression levels of proteins involved in the TAK1/NF-κB pathway were also assessed using western blot. The levels of miR-143-3p were found to be markedly lower in samples from patients with GDM, which were in turn negatively correlated with blood glucose levels. Overexpression of miR-143-3p in MIN6 cells significantly reversed HG-induced cell apoptosis and impairments in cell viability and insulin secretion. In addition, miR-143-3p overexpression attenuated HG-induced proinflammatory cytokine production by MIN6 cells. Subsequently, TGFß-activated kinase 1 (TAK1), an upstream regulator of the NF-κB pathway, was found to be a direct target of miR-143-3p in pancreatic ß cells through luciferase assays and western blot. Overexpression of TAK1 was revealed to abolish the curative effects of miR-143-3p on insulin secretion, cell viability and inflammatory response in HG-treated MIN6 cells. In addition, miR-143-3p could inactivate the NF-κB pathway by inhibiting TAK1 expression. Collectively, these results suggest that miR-143-3p levels are downregulated in the peripheral blood of patients with GDM. Therefore, miR-143-3p overexpression may serve as a method for preventing pancreatic ß cell dysfunction by inhibiting the TAK1/NF-κB pathway.

Identification and validation of an angiogenesis-related signature associated with preeclampsia by bioinformatic analysis.

Preeclampsia (PE) is a pregnancy disorder with high morbidity and mortality rates for both mothers and newborns. This study explores potential diagnostic indicators of PE. We downloaded the messenger ribonucleic acid profiles of the GSE75010 dataset from the Gene Expression Omnibus database, and used placenta samples to carry out different analyses including differential expression, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes analyses. Least absolute shrinkage and selection operator regression was constructed and the receiver operating characteristic curve was drawn to evaluate the accuracy of the model. An external validation was conducted to prove the stability of the risk model. We found 140 angiogenesis-related genes and identified 29 angiogenesis-related genes between the 2 groups, including 12 upregulated genes and 17 downregulated genes. In addition, we established a 12-gene risk signature, which has a high accuracy in predicting PE during pregnancy (area under curve = 0.90). The immune infiltration characteristics are differentially distributed in the 2 groups, which may be the cause of hypertension during pregnancy. The external validation with the GSE25906 dataset confirmed the high accuracy of our model (area under curve = 0.87). Our results outline the characteristics of a set of genes potentially involved in PE and its subgroups, contributing to a better understanding of the molecular mechanisms of PE.

Synergistic effects of thalidomide and cisplatin are mediated via the PI3K/AKT and JAK1/STAT3 signaling pathways in cervical cancer.

Thalidomide (THD) has been found to synergize with cisplatin (DDP) in certain types of cancers; however, their combined use in the treatment of cervical cancer has not been reported to date, at least to the best of our knowledge. Thus, the present study aimed to explore the synergistic effects of THD and DDP and determine their regulatory effects on the phosphoinositide 3­kinase (PI3K)/protein kinase B (AKT) and Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) pathways in cervical cancer. For this purpose, 0­160 µM THD and 0­64 µM DDP monotherapy or in combination were used to treat the HeLa and SiHa cervical cancer cell lines. This was followed by the calculation of the combination index (CI) and 160 µM THD and 16 µM DDP were then used to treat the cells. Relative cell viability and apoptosis, as well as the mRNA and protein levels of PI3K, AKT, JAK1 and STAT3 were evaluated. The results revealed that THD and DDP monotherapy suppressed the viability of the HeLa and SiHa cells in a concentration­dependent manner. Moreover, THD and DDP treatment exerted a more prominent suppressive effect on the relative viability of HeLa and SiHa cells compared with DDP monotherapy at several concentration settings; further CI calculation revealed that the optimal synergistic concentrations were 160 µM for THD and 16 µM for DDP. Subsequently, combined treatment with THD and DDP suppressed relative cell viability, whereas it promoted cell apoptosis compared with THD or DPP monotherapy; it also inhibited the PI3K/AKT and JAK1/STAT3 signaling pathways compared with DPP or THD monotherapy in both HeLa and SiHa cells. On the whole, the present study demonstrated that THD synergizes with DDP to exert suppressive effects on cervical cancer cell lines. This synergistic action also inactivated the PI3K/AKT and JAK1/STAT3 pathways. Thus, these findings suggest that the combined use of THD and DPP may have potential for use in the treatment of cervical cancer.

Construction and validation of a prognostic model based on 11 lymph node metastasis-related genes for overall survival in endometrial cancer.

BACKGROUND: Endometrial cancer (EC) is one of the most common malignant tumors in female reproductive system. The incidence of lymph node metastasis (LNM) is only about 10% in clinically suspected early-stage EC patients. Discovering prognostic models and effective biomarkers for early diagnosis is important to reduce the mortality rate. METHODS: A least absolute shrinkage and selection operator (LASSO) regression was conducted to identify the characteristic dimension decrease and distinguish porgnostic LNM related genes signature. Subsequently, a novel prognosis-related nomogram was constructed to predict overall survival (OS). Survival analysis was carried out to explore the individual prognostic significance of the risk model and key gene was validated in vitro. RESULTS: In total, 89 lymph node related genes (LRGs) were identified. Based on the LASSO Cox regression, 11 genes were selected for the development of a risk evaluation model. The Kaplan-Meier curve indicated that patients in the low-risk group had considerably better OS (p = 3.583e-08). The area under the ROC curve (AUC) of this model was 0.718 at 5 years of OS. Then, we developed an OS-associated nomogram that included the risk score and clinicopathological features. The concordance index of the nomogram was 0.769. The survival verification performed in three subgroups from the nomogram demonstrated the validity of the model. The AUC of the nomogram was 0.787 at 5 years OS. Proliferation and metastasis of HMGB3 were explored in EC cell line. External validation with 30 patients in our hospital showed that patients with low-risk scores had a longer OS (p-value = 0.03). Finally, we revealed that the most frequently mutated genes in the low-risk and high-risk groups are PTEN and TP53, respectively. CONCLUSIONS: Our results suggest that LNM plays an important role in the prognosis, and HMGB3 was potential as a biomarker for EC patients.

SIRT2 expression exhibits potential to serve as a biomarker for disease surveillance and prognosis in the management of cervical cancer patients.

This study aimed to compare the sirtuin 2 (SIRT2) expression between tumor tissue and adjacent tissue, and to investigate the association of tumor SIRT2 expression with clinical characteristics and survival profiles in cervical cancer patients.One hundred ninety-one cervical cancer patients were reviewed in this retrospective study. All patients underwent surgical resection and had well-preserved tumor tissue and adjacent tissue, which were obtained for SIRT2 expression detection by immunohistochemistry (IHC). Clinical parameters were obtained. Disease free survival (DFS) and overall survival (OS) were calculated.Both SIRT2 expression by IHC score (P < .001) and the percentage of SIRT2 high expression (defined as IHC score >3) (P < .001) were declined in tumor tissue compared with paired adjacent tissue. In addition, SIRT2 expression in tumor tissue was negatively correlated with tumor size (P = .047), lymph node metastasis (P = .009) and FIGO stage (P = .001). And the DFS (P = .007) as well as OS (P = .008) were better in patients with SIRT2 high expression compared with patents with SIRT2 low expression. Univariate Cox's proportional hazards regression model analyses revealed that high SIRT2 expression in tumor tissue was a predictive factor for more prolonged DFS (P = .009) and OS (P = .011), while multivariate Cox's proportional hazards regression model analysis disclosed that it lacks independent predictive value for DFS (P = .084) or OS (P = .132).SIRT2 expression exhibits potential to serve as a biomarker for disease surveillance and prognosis in the management of cervical cancer patients.

Cordblood-Based High-Throughput Screening for Deafness Gene of 646 Newborns in Jinan Area of China.

OBJECTIVES: Infants with slight/mild or late-onset hearing impairment might be missed in universal newborn hearing screening (UNHS). We identified the mutation hot spot of common deaf gene in the newborns in Jinan area population by screening the mutation spot with neonate cord blood, in order to make clear whether the neonate cord blood for screening is feasible. METHODS: Six hundred and forty-six newborns were subjected to both UNHS and genetic screening for deafness by using neonate cord blood. The newborn genetic screening targeted four deafness-associated genes, which were commonly found in the Chinese population including gap junction beta-2 protein (GJB2), gap junction beta-3 protein (GJB3), solute carrier family 26 member 4 (SLC26A4), and mtDNA 12S rRNA. The most common 20 spot mutations in 4 deaf genes were detected by MassARRAY iPLEX platform and mitochondrial 12S rRNA A1555G and C1494T mutations were sequenced using Sanger sequencing. RESULTS: Among the 646 newborns, 635 cases passed the UNHS and the other 11 cases (1.7%) did not. Of the 11 failures, two cases were found to carry homozygous GJB2 p.R143W pathogenic mutation, one case was found to have heterozygous GJB2 235delC mutation, and another one case carried heterozygous GJB3 p.R180X pathogenic mutation. Six hundred and thirty-five babies passed the newborn hearing screening, in which 25 babies were identified to carry pathogenic mutations, including 12 heterozygotes (1.9%) for GJB2 235delC, eight heterozygotes (1.3%) for SLC26A4 IVS7-2A>G, one heterozygote (0.2%) for p.R409H, two homozygotes (0.3%) for m.1494C>T, and two homozygotes (0.3%) for m.1555A>G. CONCLUSION: Newborn genetic screening through the umbilical cord blood for common deafness-associated mutations may identify carriers sensitive to aminoglycoside antibiotic, and can effectively prevent or delay hearing loss occurs.

[Systematic evaluation of the new screen methods of cervical intraepithelial neoplasm].

OBJECTIVE: To evaluate new cytologic screening methods of papnet cytology computer technology (CCT) and thinprep pap test (TPT) on cervical intraepithelial neoplasm (CIN). METHODS: Three hundred and seventy four cases with abnormal cervical cytologic results received colposcopic examinations and multiple biopsies. The results of cytologic screening compared with the results of histologic-colposcopic diagnosis were analysed retrospectively. RESULTS: A total of 374 cases were diagnosed by cytology, 9.1% with inflammations, 43.3% with low grade squamous intraepithelial lesion (LSIL) and 20.3% with high grade squamous intraepithelial lesion (HSIL) and 1.9% suspected with squamous cell carcinoma. However, after examination by colposcopy and multiple biopsies, the situation was different. The percentage of cases with inflammation increased to 53.5%, cervical intraepithelial neoplasm I (CIN I) decreased to 17.9%, CIN II and CIN III and cervical carcinoma in situ (CIS) elevated to 25.4% and invasive squamous cell carcinoma increased to 3.2%. Among cases with atypical squamous cell of undetermined significance (ASCUS), 52.6% (50/95) were with inflammations or negative results, 47.4% (45/95) with CIN I or even greater. Among 374 cases, 188 women were screened by CCT and 186 by TPT. When all cytological findings compared with the histologic-colposcopic results, respectively, the coincidence of TPT with histologic-colposcopic results was significantly higher than that of CCT (P < 0.01). The coincidence rate (79.7%) of screening cases with LSIL or inflammation was significantly higher than that (20.3%) of cases with HSIL or even greater (P < 0.01). But there was no difference between two methods. The incidence of human papillomaviral infection (HPV) infection at colposcopies was 34.0%. The high incidences of HPV and CIN infection were found among women from 20 to 30 years old and from 30 to 50 years old, respectively. CONCLUSIONS: Cases with abnormal cytologic findings should undergo colpscopic examination and multiple biopsies for further diagnosis. Only in this way, cases with CIN or HPV infection could not be misdiagnosed. Women ranged from 30 - 50 years old should receive cytologic screening or colposcopic examination regularly.

Combination of hearing screening and genetic screening for deafness-susceptibility genes in newborns.

The aim of this study was to determine the clinical significance of the results of screening of newborn hearing and the incidence of deafness-susceptibility genes. One thousand newborn babies in the Handan Center Hospital (Handan, China) underwent screening of hearing and deafness-susceptibility genes. The first screening test was carried out using otoacoustic emissions (OAEs). Babies with hearing loss who failed to pass the initial screening were scheduled for rescreening at 42 days after birth. Cord blood was used for the screening of deafness-susceptibility genes, namely the GJB2, SLC26A4 and mitochondrial 12S rRNA (MTRNR1) genes. Among the 1,000 neonates that underwent the first hearing screening, 25 exhibited left-sided hearing loss, 21 exhibited right-sided hearing loss and 15 cases had binaural hearing loss. After rescreening 42 days later, only one of the initial 61 cases exhibited hearing loss under OAE testing. The neonatal deafness gene tests showed two cases with 1555A>G mutation and two cases with 1494C>T mutation of the MTRNR1 gene. In the SLC26A4 gene screening, four cases exhibited the heterozygous IVS7-2A>G mutation and one case exhibited heterozygous 1226G>A mutation. In the GJB2 gene screening, two cases exhibited the homozygous 427C>T mutation and 10 exhibited the heterozygous 235delC mutation. The genetic screening revealed 21 newborns with mutations in the three deafness-susceptibility genes. The overall carrier rate was 2.1% (21/1,000). The association of hearing and gene screening may be the promising screening strategy for the diagnosis of hearing loss.