Cervical cancer is a serious disease endangering the health of women, which is next only to breast cancer and colon cancer.There are nearly 130 000 new cases of cervical cancer arise in China each year, accounting for 1/5 of the total number of new cases in the world.Epidemiological survey has found that cervical cancer is related to various factors, such as human papilloma virus infection, multiple sexual partners, smoking, premature sexual life, sexually transmitted diseases, economic status and immunosuppression.Cervical cancer is pathologically characterized by cervical dysplasia cervical atypical hyperplasia (mild, moderate, severe) to carcinoma in situ and to infiltrating carcinoma.Female vagina is a relatively anoxic and unique dynamic micro ecological system colonized with a large number of bacteria.And the vaginal microflora is the kernel of maintaining vaginal micro ecological balance.The uterine cervix is directly exposed to the vagina, and is closely related to vaginal microbes.However, there are few reports on the direct relationship between cervical cancer and vaginal microbes.In this review, the roles and significance of vaginal microbial imbalance and common vaginal infections in cervical cancer will be discussed from different angles.

ZnO nanoparticle-containing carbon composite nanofiber ( ZnO-CNF ) was prepared by the electrospinning of the ZnCl2-PAN precursor, followed by preoxidation and carbonization. The ZnO nanoparticles were uniformly distributed on the surface of the carbon nanofiber with the size of 20-30 nm, confirmed by scanning electron microscopy ( SEM ) . The wettability of the ZnO-CNF was studied by water contact angle test. With Nafion as an additive, the ZnO-CNF modified electrode was successfully constructed by dip-coating. The surface morphology and electrochemical properties of the modified electrode were investigated by SEM and cyclic voltammetry. There was a sensitive response of the ZnO-CNF modified electrode on Pb ions in solution, demonstrated by square wave stripping voltammetry. Under the optimized conditions, a good linear relationship between peak current and Pb2+concentration was obtained in the range of 2. 4×10-10-2. 4×10-7 mol/L (R=0. 998) by 10 min preconcentration at -1. 0 V in 0. 1 mol/L NaAc buffer solution (pH=4. 6). The detection limit was 4. 8×10-11 mol/L. The practical analytical application of the ZnO-CNF modified electrode was assessed by the measurement of the actual water sample and the result was consistent with that obtained by ICP-MS.

OBJECTIVETo explore the role of HIF1α gene in prostate cancer cell line DU145 by knocking it out with a novel gene-editing tool CRISPR/cas9 system.

METHODSA CRISPR/cas9 system with two sgRNAs targeting exon 1 of the HIF1α gene was constructed for the knock out experiment. CCK8 assay and transwell experiment were carried out to assess the effect of the knock out on the proliferation, migration and invasiveness of DU145 cells.

RESULTSThe efficiency of gene-targeting was measured through a T7E1 assaying and sequence analysis, which confirmed that the partial knock out was successful and has led to a significant decrease in the expression of HIF1α and inhibition of cell proliferation, migration and invasiveness.

CONCLUSIONA CRISPR/cas9 system for the knock out of HIF1α has been successfully constructed, which could inhibit the proliferation and migration of DU145 cells. The system can facilitate further studies of the HIF1α gene and its roles in tumorigenesis.

CRISPR-Cas Systems ; genetics ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Editing ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; physiology ; Male ; Neoplasm Invasiveness ; Prostatic Neoplasms ; pathology

Objective: To investigate the status and prognostic significance of TERT and IDH1/2 genes mutations in diffusely infiltrating gliomas. Methods: Hot spot mutations of TERT and IDH1/2 genes were detected by DNA sequencing in 236 cases of gliomas at West China Hospital from 2012 to 2016, including pilocytic astrocytoma (WHO grade Ⅰ, 16 cases), diffuse astrocytoma and oligodendroglioma (WHO grade Ⅱ, 89 cases), anaplastic astrocytoma and oligodendroglioma (WHO grade Ⅲ, 72 cases) and glioblastoma (WHO grade Ⅳ, 59 cases). The prognostic significance of TERT and IDH1/2 hot spot mutations was evaluated. Results: No IDH or TERT mutations were detected in pilocytic gliomas. TERT promoter mutation frequency was higher in patients aged ≥40 years(60.8%, 93/153) than in patients aged <40 years (32.8%, 22/67; P<0.01). TERT promoter mutation rate was also significantly higher in oligodendroglioma (87.5% , 56/64) than that in astrocytoma(37.8%, 59/156; P<0.01). Young age (<40 years), oligodendroglioma and IDH1 mutation were favorable prognostic factors for diffusely infiltrating astrocytic and oligodendroglial tumors. TERT mutation alone was not of prognostic significance. Diffusely infiltrating astrocytic and oligodendroglial tumors were divided into four molecular subtypes according to TERT and IDH1 mutation status: IDH(+ )/TERT(+ ), IDH(+ )/TERT(-), IDH(-)/TERT(-) and IDH(-)/TERT(+ ). There was significant prognostic difference among the 4 subtypes. Conclusions Combined IDH and TERT gene mutation analysis may be useful for prognostic subgrouping. Notably, IDH1 wild-type cases can be further subdivided into TERT(+ ) or (-) subgroups with significant prognostic difference.

Objective:To investigate the clinicopathological and molecular characteristics of the epithelioid glioblastoma (eGBM) with BRAF V600E mutation.Methods:Sixteen cases of eGBM with BRAF V600E mutation diagnosed at the West China Hospital of Sichuan University, China from 2012 to 2019 were collected. Their clinicopathological and molecular characteristics were analyzed. Results:The range of patients′ age was from 7 to 61 years (median 31.5 years). There were 4 males and 12 females, with a male to female ratio of 1∶3. Eleven cases were newly diagnosed eGBM and five cases had a previous history of astrocytomas. Most of the tumors were located in the cerebral hemisphere, often in the frontal lobe, with an average diameter of 4.6 cm (2.0-8.0 cm). The tumors were composed of relatively uniform, closely packed epithelioid cells, some showing discohesion, with distinct cell membrane, eosinophilic cytoplasm, eccentric nuclei, distinct nucleoli and mitotic activity. Palisaded/coagulative necrosis was seen in all cases. Glomerular microvascular proliferation was seen in most of the cases, while mono-or multi-nucleated tumor giant cells were seen in some cases. Focal sarcomatoid area was seen in 2 cases, and focal pleomorphic xanthoastrocytoma (PXA)-like area was seen in 3 cases. Immunohistochemistry showed variable positivity for GFAP, Olig2 and p53. The median Ki-67 index was 30% (10%-50%). Only one case lost ATRX protein expression. Sanger sequencing identified the BRAF V600E mutation in all sixteen patients. Five cases also had mutations in the TERT gene promoter. No IDH1 (R132) or IDH2 (R172) mutation was detected. Surgical resection of the tumors was performed for all patients, and 3 patients also received adjuvant radiotherapy and chemotherapy. Follow-up data were available for 15 patients, with a follow-up time of 1-89 months (median 10 months). Among the 15 patients, 7 patients died of disease and another 5 patients had recurrences. The overall survival time of the patients under 35 years of age was significantly longer than that of the patients aged 35 years or older ( P=0.014), but their progression-free survival was not statistically different ( P=0.232). Conclusions:eGBM with BRAF V600E mutation is more commonly detected in young women than other the populations (i.e. elderly or male). The epithelioid morphology should include rhabdoid meningioma, anaplastic PXA, atypical teratoid/rhabdoid tumor, metastatic tumors, and melanoma in its differential diagnosis. PXA-like area is observed in some eGBM cases, suggesting a relationship of these two types of tumor. eGBM is a high-grade malignant tumor and most of the cases show recurrences or deaths in a short-period time. The younger patients have a relatively better prognosis than the older ones.

Objective:To investigate the clinical and pathological features and prognosis of patients with microfocal prostate adenocarcinoma.Methods:Clinical and pathological data of the patients diagnosed with microfocal adenocarcinoma on prostate biopsy at the West China Hospital from 2013 to 2019 were collected. Microfocal adenocarcinoma was defined as follows: Gleason score of 3+3=6, total number of the cores ≥10, number of the positive cores ≤2, and proportion of the tumor in each positive core<50%. Clinicopathological parameters, treatment plans and follow-up data were collected. Pathological information of the biopsy and radical resection specimens was used to analyze the correlation between pathological parameters in the biopsy report and adverse pathological features of radical resection specimens, including increased Gleason score, capsule invasion, positive surgical margin and perineural invasion.Results:A total of 206 cases of microfocal adenocarcinoma were diagnosed on prostate biopsies from 2013 to 2019, accounting for 6.7% of all adenocarcinoma cases. There were 139 cases of 1 positive core and 67 cases of 2 positive cores. Patients with microfocal adenocarcinoma were younger than those with non-microfocal adenocarcinoma (69 years versus 71 years, P<0.001). Compared with patients with non-microfocal adenocarcinoma, the pre-biopsy total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) levels in patients with microfocal adenocarcinoma were both lower (11.2 μg/L 2 versus 23.7 μg/L 2; 1.4 μg/L 2 versus 3.0 μg/L 2, P<0.001), the fPSA/tPSA level was higher (12.9% versus 10.7%, P<0.05), the prostate volume was larger (38.9 mL versus 34.3 mL, P<0.05), and the PSA density was lower (0.3 μg/L 2 versus 0.8 μg/L 2, P<0.001). 130 patients underwent radical prostatectomy, 30 patients chose active monitoring, 31 patients chose endocrine or radiation therapy, and 15 patients were lost to follow-up. Three patients in the active surveillance group underwent radical prostatectomy for disease progression after 21-39 months observation. Biochemical relapses occurred in two patients in the radical prostatectomy group. The remaining patients have no disease progression or recurrence at present. Compared with radical prostatectomy specimens, Gleason score in the biopsy material was increased in 64/115 patients (55.7%). Among resection excision specimens, 14 cases (12.2%) had extraprostatic extension (EPE), 35 cases (30.4%) had perineural invasion, and 16 cases (13.9%) had a positive margin. Univariate and multivariate analyses showed that low fPSA/tPSA ratio and 2 positive cores were independent risk factors for Gleason score increase in the radical prostatectomy specimens. A low fPSA/tPSA ratio was an independent risk factor for perineural invasion. Low fPSA/tPSA ratio and low prostate volume were associated with a positive margin in radical prostatectomy specimens. Conclusions:In this study, patients diagnosed with microfocal adenocarcinoma on prostate biopsy account for a high proportion of the patients with increased Gleason score in the radical prostatectomy specimens, and there is a certain proportion of adverse pathological features in the radical specimens. Therefore, for the patients with only a small amount of low-grade adenocarcinoma found in biopsy, PSA levels and PSA density should be taken into consideration in treatment selection.

Objective:To study the clinicopathological features, immunophenotype, molecular genetic characteristics and prognosis of renal cell carcinoma associated with TFEB gene rearrangement (TFEBr-RCC).Methods:Eight cases of TFEBr-RCC diagnosed at the West China Hospital of Sichuan University from 2014 to 2022 were collected for clinicopathological, immunohistochemical, fluorescence in situ hybridization and RNA sequencing analyses, with review of literature.Results:Six patients were male and two were female. The patient ages ranged from 25 to 50 years (mean: 34 years, median: 32 years). The tumors were present in the right kidney (3 cases) or the left kidney (5 cases). The maximum diameters of the tumors ranged from 4.0 cm to 18.5 cm, with an average of 8.5 cm. Histologically, majority of the cases (5/8) showed typical biphasic "pseudorosette" structure, while the remaining three cases demonstrated atypical morphology that was similar to epithelioid angiomyolipoma or clear cell renal cell carcinoma. Immunohistochemical study showed positivity of TFEB (8/8), PAX8 (8/8), MART-1 (7/7), and HMB45 (5/6). Interestingly, PD-L1 was variably expressed in all five tested cases. Staining for TFE3 in all cases was negative. TFEB translocation was verified in all 8 cases using TFEB fluorescence in situ hybridization. RNA sequencing showed MALAT1-TFEB gene fusion in 4 of the 5 tested cases (two of which showing novel MALAT1-TFEB fusion sites), and one case with a novel ACTB-TFEB gene fusion. Patient follow-ups ranged from 5 to 96 months (average 47 months). All patients were alive without recurrence or metastasis.Conclusions:TFEBr-RCC tends to occur in young adults and has a good prognosis. Histologically, most of the cases show characteristic biphasic structure, and some cases show epithelioid angiomyolipoma-like or clear cell RCC-like morphology. Immunohistochemical reactivity to TFEB, melanocytic markers and PD-L1 is characteristic. MALAT1-TFEB gene fusion is the most common molecular change, with variable fusion sites.

Objective:To investigate the expression of HER2 and its relationship with clinicopathological features in patients with urothelial carcinoma.Methods:Urothelial carcinoma specimens collected from January 2019 to June 2022 were used. The expression of HER2, cytokeratin 20 and cytokeratin 5/6 was examined using immunohistochemistry. The HER2 expression was assessed according to the clinical pathological expert consensus on HER2 testing in urothelial carcinoma in China. Cases with HER2 2+/3+ were classified as HER2 positive. The relationship between HER2 expression and clinicopathological and molecular features was analyzed.Results:Four hundred and twenty-nine urothelial carcinoma specimens were analyzed, including 166 cases of raclical resection and 263 cases of local tumor resection. The median patient-age was 69 years (range: 31-93 years). The male: female ratio was 2.9∶1.0. The positive rate of HER2 was 45.7%(196/429). The positive rate of HER2 in patients with local tumor resection was higher than that in patients with radical resection [51.7%(136/263) vs.36.1%(60/166), P<0.05]. In the upper urinary tract (renal pelvis/ureter) urothelial carcinomas, the positive rate of HER2 was 35.2% (37/105). In bladder urothelial carcinoma, the positive rate of HER2 was 49.1%(157/320), and higher than that in upper urinary tract urothelial carcinoma ( P<0.05). In high grade urothelial carcinoma, the positive rate of HER2 was 52.8%(168/318) and higher than that in low grade urothelial carcinomas (25.2%, 28/111, P<0.01). In 166 radical resection specimens, the positive rate of HER2 was not differentially distributed by tumor pT stage [Ta (26.1%, 6/23), T1 (41.7%, 20/48), T2 (40.0%, 10/25), T3 (28.1%, 16/57), T4 (8/13) ( P>0.05)]. In urothelial carcinomas with muscle invasion, the HER2 positive rate was 35.8%(34/95), while the rate in non-muscle-invasion urothelial carcinoma was 36.6%(26/71, P>0.05). CK20 and CK5/6 were used to refine the urothelial carcinoma molecular subtypes. The positive rate of HER2 was highest in CK20 +/CK5/6 -group (124/194, 63.9%), followed by CK20 +/CK5/6 +group (18/40, 45.0%), CK20 -/CK5/6 -group (14/41, 34.1%) and CK20 -/CK5/6 +group (25/131, 19.1%, P<0.01). The positive rate of HER2 in micropapillary urothelial carcinoma was highest (14/15), followed by urothelial carcinoma with glandular differentiation (11/14), conventional urothelial carcinoma (161/360, 44.7%) and urothelial carcinoma with squamous differentiation (6/35, 17.1%, P<0.01). In the cases with lymph node metastasis, the positive rate of HER2 was 45.5% (10/22) and higher than the cases without lymph node metastasis (31.0%, 13/42). But there was no statistically significant association between HER2 expression and lymph node metastasis ( P>0.05). Conclusions:Expression of HER2 in urothelial carcinoma is closely correlated with tumor location, grade, histologic subtypes, molecular subtypes and surgical approach, but not with pT stage, muscle invasiveness or lymph node metastasis.

Objective:To analyze the clinicopathological features of prostate cancers with BRCA2 pathogenic mutations, and the association between BRCA2 pathogenic mutation and clinicopathological characteristics. Patient survivals were also examined.Methods:Clinicopathological data of 249 prostate cancer patients who underwent genetic testing in West China Hospital of Sichuan University, Chengdu, China from June 2014 to August 2021 were collected. A retrospective analysis of histopathological morphology, clinicopathological characteristics, and patient survivals was conducted.Results:The genetic testing in the 249 prostate cancer patients showed a pathogenic mutation of DNA damage repair gene (DRG) in 73 cases (73/249, 29.3%), including 22 cases (8.8%) with BRCA2 pathogenic mutation and 51 cases with pathogenic mutations of other DRG. Among the 22 patients with BRCA2 pathogenic mutation, 14 patients (5.6%) harbored germline mutations and 8 patients (3.2%) somatic mutations. Their ages ranged from 48 to 91 years, with a median of 67 years. Seventeen patients (77.3%) had distant metastasis, including 16 cases with bone metastasis and 1 case with multiple metastases. Thirteen patients (59.1%) were castration-resistant prostate cancer. The histological type was mainly classical prostatic acinar adenocarcinoma, including 16 cases (72.7%) with intraductal carcinoma of the prostate (IDC-P). Six cases (27.3%) showed focal neuroendocrine differentiation. Perineural/vascular invasion and extraprostatic extension were seen in 11 cases (50.0%) and 8 cases (36.4%), respectively. The Gleason scores of 19 patients (86.4%) were≥8. IDC-P was more commonly found in patients with BRCA2 germline pathogenic mutation than those with BRCA2 somatic pathogenic mutation, other DRG pathogenic mutation or no-DRG pathogenic mutation ( P=0.002). With a total follow-up time of 189 months, the median overall survival (OS) was 132.3 months. Patients with DRG pathogenic mutation had shorter OS than those with no-DRG pathogenic mutation ( P=0.040). The OS of patients with BRCA2 germline pathogenic mutation did not significantly differ from that of patients with BRCA2 somatic pathogenic mutation, other DRG pathogenic mutation or no-DRG pathogenic mutation ( P=0.216). Conclusions:The presence of BRCA2 gene pathogenic mutation is common in the prostate cancers with high Gleason grade, advanced clinical stage, and castration resistance. IDC-P is more commonly noted in cases with BRCA2 germline pathogenic mutation than those without. Patients with DRG pathogenic mutation have shorter OS than those with no-DRG pathogenic mutation, but there is no significant association between BRCA2 pathogenic mutations and OS.

OBJECTIVE: The medicinal mushroom Sanghuangporus vaninii produces pharmaceutically valuable hispidin polyphenols in natural habitats. However, due to the slow growth in nature, S. vaninii grown in the field (sclerotia) is not reliable for pharmaceutical purposes. Although higher biomass of fungal mycelia can be obtained in submerged cultures, the accumulation of hispidin polyphenols is rare. METHODS: In this study, the polyunsaturated fatty acids (PUFAs), linoleic acid (LA), linolenic acid (ALA), and methyl jasmonate (MeJa) were employed as the stimulant agents to coordinate the accumulation of biomass and hispidin polyphenols in its submerged cultures. RESULTS: The addition of LA and ALA promoted the mycelial accumulation, while the addition of MeJa inhibited the growth of S. vaninii concomitant with reduced total polyphenols. UPLC-Triple-TOF-MS analysis revealed an increased production of hispidin, phellinstatin, pinnilidine, and its derivatives upon the addition of LA and ALA, and hypholomine B and its isomer, 3,14'-bihispidinyl, and phelligridin E upon the addition of MeJa on day 13. Intriguingly, total polyphenols from the MeJa-supplementing cultures harbored a high capacity in scavenging free radicals. Chemical structural analysis showed that hispidin polyphenols had higher antioxidant activity due to more hispidin moieties induced by MeJa. CONCLUSION The supplement of PUFAs affects the synthesis and composition of hispidin polyphenols in S. vaninii. Our results provide a possibility to coordinate the production of hispidin polyphenols via submerged cultures of S. vaninii.