[The expression and function of PD-L1 in CD133(+) human liver cancer stem-like cells].

Objective: To investigate the expression of programmed death protein-ligand 1 (PD-L1) in liver cancer stem-like cells (LCSLC) and its effect on the characteristics of tumor stem cells and tumor biological function, to explore the upstream signaling pathway regulating PD-L1 expression in LCSLC and the downstream molecular mechanism of PD-L1 regulating stem cell characteristics, also tumor biological functions. Methods: HepG2 was cultured by sphere-formating method to obtain LCSLC. The expressions of CD133 and other stemness markers were detected by flow cytometry, western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the expressions of stemness markers and PD-L1. The biological functions of the LCSLC were tested by cell function assays, to confirm that the LCSLC has the characteristics of tumor stem cells. LCSLC was treated with cell signaling pathway inhibitors to identify relevant upstream signaling pathways mediating PD-L1 expression changes. The expression of PD-L1 in LCSLC was down regulated by small interfering RNA (siRNA), the expression of stem cell markers, tumor biological functions of LCSLC, and the changes of cell signaling pathways were detected. Results: Compared with HepG2 cells, the expression rate of CD133 in LCSLC was upregulated [(92.78±6.91)% and (1.40±1.77)%, P<0.001], the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were also higher than those in HepG2 cells (P<0.05), the number of sphere-formating cells increased on day 7 [(395.30±54.05) and (124.70±19.30), P=0.001], cell migration rate increased [(35.41±6.78)% and (10.89±4.34)%, P=0.006], the number of transmembrane cells increased [(75.77±10.85) and (20.00±7.94), P=0.002], the number of cloned cells increased [(120.00±29.51) and (62.67±16.77), P=0.043]. Cell cycle experiments showed that LCSLC had significantly more cells in the G(0)/G(1) phase than those in HepG2 [(54.89±3.27) and (32.36±1.50), P<0.001]. The tumor formation experiment of mice showed that the weight of transplanted tumor in LCSLC group was (1.32±0.17)g, the volume is (1 779.0±200.2) mm(3), were higher than those of HepG2 cell [(0.31±0.06)g and (645.6±154.9)mm(3), P<0.001]. The expression level of PD-L1 protein in LCSLC was 1.88±0.52 and mRNA expression level was 2.53±0.62, both of which were higher than those of HepG2 cells (P<0.05). The expression levels of phosphorylation signal transduction and transcription activation factor 3 (p-STAT3) and p-Akt in LCSLC were higher than those in HepG2 cells (P<0.05). After the expression of p-STAT3 and p-Akt was down-regulated by inhibitor treatment, the expression of PD-L1 was also down-regulated (P<0.05). In contrast, the expression level of phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in LCSLC was lower than that in HepG2 cells (P<0.01), there was no significant change in PD-L1 expression after down-regulated by inhibitor treatment (P>0.05). After the expression of PD-L1 was knockdown by siRNA, the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were decreased compared with those of siRNA-negative control (NC) group (P<0.05). The number of sphere-formating cells decreased [(45.33±12.01) and (282.00±29.21), P<0.001], the cell migration rate was lower than that in siRNA-NC group [(20.86±2.74)% and (46.73±15.43)%, P=0.046], the number of transmembrane cells decreased [(39.67±1.53) and (102.70±11.59), P=0.001], the number of cloned cells decreased [(57.67±14.57) and (120.70±15.04), P=0.007], the number of cells in G(0)/G(1) phase decreased [(37.68±2.51) and (57.27±0.92), P<0.001], the number of cells in S phase was more than that in siRNA-NC group [(30.78±0.52) and (15.52±0.83), P<0.001]. Tumor formation in mice showed that the tumor weight of shRNA-PD-L1 group was (0.47±0.12)g, the volume is (761.3±221.4)mm(3), were lower than those of shRNA-NC group [(1.57±0.45)g and (1 829.0±218.3)mm(3), P<0.001]. Meanwhile, the expression levels of p-STAT3 and p-Akt in siRNA-PD-L1 group were decreased (P<0.05), while the expression levels of p-ERK1/2 and ß-catenin did not change significantly (P>0.05). Conclusion: Elevated PD-L1 expression in CD133(+) LCSLC is crucial to maintain stemness and promotes the tumor biological function of LCSLC.

Correlation between Obstructive Sleep Apnea and Non-Alcoholic Fatty Liver Disease before and after Metabolic Bariatric Surgery.

BACKGROUND: Emerging evidence has revealed that obstructive sleep apnea (OSA) is associated with non-alcoholic fatty liver disease (NAFLD). However, the impact of OSA on NAFLD among obese patients undergoing metabolic and bariatric surgery (MBS), especially during follow-up period, remains unclear. OBJECTIVE: To analyze the correlation based on preoperative characteristics and postoperative conditions among bariatric patients with comorbid OSA and NAFLD. METHODS: Clinical data of patients who underwent MBS in our institution between January 2016 and June 2019 were reviewed retrospectively. Correlation analysis and linear regressions were used to identify how OSA links with NAFLD before and after treatment of MBS. RESULTS: Of 308 patients, 181 were diagnosed with OSA and enrolled in the present study, and 127 completed follow-up visits at 6 months. The proportion of NAFLD in the mild-moderate OSA and severe OSA groups was 75.0% and 96.0%, respectively. MBS was effective at improving sleep apnea and nocturnal hypoxia, as well as liver steatosis and fibrosis (P < 0.05). And we also found that there were significant correlations not only between OSA- and NAFLD-related characteristics at baseline but also between their improvements after surgery, eventually leading to similar prognosis of NAFLD for both groups (P < 0.05), no matter what presurgical differences existed. In addition, the results of the univariate and multivariate linear regression analyses supported preoperative liver/spleen Hounsfield units ratio (LSR) by computerized tomography (CT) as an independent predictor of the effect of MBS on liver steatosis. CONCLUSION: In conclusion, MBS plays a pivotal role in the control of medical conditions in obese patients with OSA and NAFLD. Given the correlation between OSA and NAFLD in the present study, in the case of both the severity at baseline as well as the improvement after surgery, OSA may pose an impact on the prognosis of NAFLD in bariatric patients.

MiR-491 suppresses migration and invasion via directly targeting TPX2 in breast cancer.

OBJECTIVE: Breast cancer (BC) is one of the primary causes of tumor-related female mortalities. Although in recent years, we have made great progress in the systemic therapy and earlier diagnosis for BC patients, recurrence or distant metastasis remains leading obstacles for the successful therapy of BC. Therefore, a comprehensive understanding of the molecular mechanism underlying the progression may be crucial in developing an effective strategy against BC. The current research aimed to explore the expressions, functions and molecular mechanism of microRNA-491 (miR-491) in BC. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the level of miR-491 expression in 52 pairs of BC tissues and para-cancerous specimens, and the relation between miR-491 level and the clinical features of BC patient prognosis was analyzed. Transwell invasion and migration assays were conducted to determine whether miR-491 had effects on the regulation of BC metastasis. Potential target genes of miR-491 were found out using TargetScan to explore the molecular functions of miR-491 in inhibiting breast cancer cell invasion and migration. To elucidate the mechanism of TPX2 in suppressing cell invasion and migration medicated by miR-491in breast cancer, we further transfected TPX2 siRNAs into MCF-7 cells to delete endogenous TPX2, along with the transfections with miR-491 inhibitor into MCF-7 cell lines. RESULTS: The findings demonstrated that miR-491 expressions were significantly decreased in BC tissues and cells. The miR-491 restoration suppressed the invasion and migration of BC cells. In addition, we identified the targeting protein for Xklp2 (TPX2) as a direct target of miR-491 in BC. The knockdown of TPX2 markedly reversed miR-491-medicated inhibition of cell invasion and migration in BC cell lines. CONCLUSIONS: In short, all the results suggested that miR-491 functioned as a tumor suppressor by targeting TPX2 in BC and the miR-491 restoration may be an effective therapy for the BC treatment in the future.

miR-129b suppresses cell proliferation in the human lung cancer cell lines A549 and H1299.

Lung cancer is one of the most prevalent malignant tumors, and is one of the primary causes of cancer-associated deaths. In 2002, an estimated 1.18 million lung cancer-associated deaths were recorded, accounting for 18% of cancer-related deaths and 2% of total mortality. Despite the great progress that has been made in lung cancer therapies, the mechanisms underlying lung cancer formation and development remain largely unknown. Meanwhile, the microRNA miR-129 has been shown to be involved in the formation of many types of cancer. Therefore, this study aims to investigate whether miR129b could suppress proliferation of lung cancer cell lines. NSCLC tissue samples were collected from the Department of Respiratory Medicine between April 2013 and December 2015. Ten normal health individuals were recruited as controls. Lung cancer cell lines A549 and H1299 were used to examine the suppressive effects of miR129b. Quantitative real-time PCR was used to detect miR129b expression. The MTT assay was used to analyze cell proliferation. Results indicated that miR-129b is down-regulated in lung cancer cell lines and NSCLC tissues. Furthermore, overexpression of miR-129b inhibited proliferation of lung cancer cells. In conclusion, miR-129b suppresses lung cancer cell proliferation, and can be a potential therapeutic target for treatment of lung cancers.

[ZNF143 is involved in CTCF-mediated chromatin interactions by cooperation with cohesin and other partners].

ZNF143 is a ubiquitously expressed transcription factor conserved in vertebrates and might regulate the expression of numerous genes. But its function in mediating chromatin interactions remains elusive. By integrated analysis of public datasets, we provided evidence that a majority of ZNF143 binding sites (BSs) were involved in CTCF-mediated chromatin interaction networks (CTCF-CINs) by overlapping with cohesin-BSs and CTCF-BSs. We further showed that only a very few CTCF-CINs were associated with ZNF143 alone, whereas those associated with ZNF143 and cohesin simultaneously were highly overlapped with constitutive, conserved CTCF-BSs and enriched at boundaries of chromatin topologically associating domains. These observations implicate that as an important partner of CTCF, ZNF143 helps it establish the conserved chromatin structure by cooperating with cohesin.

Sequence analysis of CP and HC-Pro genes of Turnip mosaic virus isolates from China.

Fourteen isolates of Turnip mosaic virus (TuMV) were obtained from the leaves of diseased cruciferous plants in China. According host tests, the isolates were classified into B-host and BR-host group. The nucleotide sequences of the coat protein (CP) and helper component proteinase (HC-Pro) genes of the isolates were determined. The CP genes consisted of 864 nucleotides encoding a polypeptide of 288 amino acids. The HC-Pro genes comprised 1374 nucleotides encoding a polypeptide of 458 amino acids. The genes CP and HC-Pro of the 14 isolates shared nucleotide sequence identities ranging from 89.2 to 99.5% and 79.1 to 99.9%, respectively. Amino acid sequence identities of CP and HC-Pro proteins ranged from 95.1 to 100% and 94.8 to 99.8%, respectively. Phylogenetic tree based on the CP gene indicated that 13 of the 14 TuMV isolates belonged to the world-B group, while the remaining isolate ZJ1 belonged to the basal-BR group. The phylogenetic tree based on the HC-Pro gene was similar to that of CP gene with the exception of the isolate JX that clustered with the Asian-BR group. Our results were consistent with the previous results demonstrating that a majority of the isolates collected from Brassica spp. belonged to the world-B group.

Plasminogen activator inhibitor-1 in brain tumors: relation to malignancy and necrosis.

Tumor necrosis is a common feature of malignant neoplasms. The pathogenesis of tumor necrosis remains poorly documented. Recent evidence has shown a correlation between the presence of tumor necrosis and low content of tissue plasminogen activator in brain tumors and significantly higher levels of plasminogen activator inhibitor-1 (PAI-1) in human glioblastomas. We subjected fresh brain tumor tissue samples (n = 197) to an enzyme-linked immunosorbent assay to determine PAI-1 content. The results were correlated with the presence of edma and necrosis on imaging studies. The samples studied were from normal brain (n = 10), low-grade gliomas (n = 26), meningiomas (n = 47), acoustic neuromas (n = 18), glioblastomas (n = 45), metastases (n = 45), and areas of tumor necrosis (n = 6). The benign tumor samples (n = 96) had 3.5 times less PAI-1 than did the malignant tumors (n = 101). Tumor necrosis samples contained 3.8 times more PAI-1 than did the nonnecrotic malignant tumor samples (P < 0.000001). The benign meningioma samples showed a similar ratio compared with their malignant counterparts (0.35 versus 1.59 ng/mg, respectively, P = 0.0004). Regression analysis results showed a strong correlation between PAI-1 and necrosis (r = 0.47, P < 0.0000028) and, to a lesser extent, brain edema (r = 0.26, P = 0.001). A negative correlation between PAI-1 and tissue plasminogen activator levels almost reached statistical significance (P = 0.07). There was no correlation between PAI-1 content and the tumor size, duration of symptoms, or the sex or age of the patients. The results of this study indicate that malignant transformation is associated with a significant increase in PAlI1 and that PAI-1 may play an integral role in the pathogenesis of tissue necrosis, perhaps via the inhibition of tissue plasminogen activator and the promotion of microthrombosis.

[Computed tomography of the retroperitoneum following nephrectomy for renal cell carcinoma].

CT of the retroperitoneum in 70 patients (98 scans) who had undergone previous nephrectomy for renal cell carcinoma was reviewed. The interval between surgical operation and CT scanning were 15 days-7 years. In 59 cases (84%), CT scans were performed within the first 2 years. After nephrectomy, the emptied renal fossae were filled with adjacent organs, such as colon (27), small intestine (21), liver (14), inferior vena cava (3), adrenal gland (2) on the right side, and spleen (28) small intestine (33), colon (15), tail of pancreas (11), adrenal gland (2) on the left. In 47 cases, where CT scans were done within 4 months after operation, there were abscess/hematoma (8) and soft tissue thickening caused by post-operative edema and or inflammation (6). Most of the early post-operative complications gradually reduced in size in the follow-up scans. Local recurrences or metastases were detected by CT scan in 15 cases, with recurrence at the excision site (5), renal bed (6), adrenal gland (4), retroperitoneal lymph nodes (7) and contralateral kidney (1). 7 patients had solitary lesions and 8 patients had multiple recurrences. In 2 patients, local recurrences were detected by routine follow-up CT scanning. In 4 cases, recurrences were proved by surgery, and the others by clinical follow-up. Local recurrences were assessed within 2 years in 10/15 cases (67%), and 4 cases within 6 months. 9 cases had venous thrombosis (IVC 9, renal vein 3), the size of which was reduced after radiotherapy in 1 case. CT is the most preferred imaging technique in evaluating post-nephrectomy patients for renal cell carcinoma.

[Unusual imaging findings of renal angiomyolipoma with giant pseudoaneurysm and intrarenal perirenal hemorrhage].

Two patients with unusual findings of renal angiomyolipoma (AML) were reported. One had intratumoral giant pseudoaneurysm with mural thrombus and hemorrhage, the other had intrarenal and perirenal hemorrhage, due to vascular malformation, rupture of small aneurysms and pseudoaneurysms within the tumor. Since imaging techniques such as CT, ultrasonography, MRI are being so commonly used, the detectability of renal AML is increased. Understanding of the various composition of renal AML and their accompanied pathology is most helpful in improving the accuracy of imaging diagnoses.

[Comparison between actions of shiquandabu pills and shiquandabu liquor on immunological parameters in mice].

The comparison shows that both Shiquandabu pills and Shiquandabu liquor are efficacious in strengthening the immunological functions of mice. No significant difference has been found between the two forms of preparation. There exists a good dose-efficacy relationship for the pills in immunological functions.

Biological significance of tissue plasminogen activator content in brain tumors.

Fresh brain-tumor samples were obtained at operation and analyzed for their content of tissue type plasminogen activator (tPA) using an activity assay (gel chromatography zymogram) and an enzyme-linked immunospecific assay. The specimens were taken from 23 glioblastomas, 35 metastatic tumors, 42 meningiomas, 16 low-grade gliomas, and seven acoustic neurinomas; seven specimens were from normal brain. A strong correlation was found between the results of the two assays (r = 0.77, p less than 0.0001). There was a threefold difference in the tPA content of the benign tumors as compared to malignant tumors (p = 0.0006), the latter having less tPA. Histologically benign meningiomas contained higher tPA than malignant meningiomas (p = 0.01); however, the difference between low-grade gliomas and high-grade gliomas was less evident. Overall regression analysis data have shown an inverse relationship between the tissue content in tPA and the presence and degree of tumor necrosis and peritumoral brain edema (p = 0.004 and p = 0.0004, respectively). This finding was most consistent in the glioblastoma group where the correlation coefficient values were r = 0.53 and r = -0.55, respectively. There was no significant correlation between the tissue tPA content and the age and sex, steroid use, or plasma tPA of the patients or the duration of symptoms. In summary, this is the first demonstration of tPA in a large series of human brain tumors and in normal brain. The differences observed have clear biological significance and, although the source of tPA in tumor tissue is still unknown, a relative reduction in tPA in tumor tissue may play an integral role in the development of tissue necrosis and tissue edema. The lack of tPA in tumor necrosis was not due to tissue destruction and cell death since urokinase was readily detectable in that tissue.

[Image diagnosis of small renal carcinoma--a report of 7 cases].

Seven cases (8 tumors) of small renal carcinoma (less than 3 cm in diameter) proven by pathology were reviewed. It accounted for 7% of patients with renal cell carcinoma during the same period. Sonogram was hypoechogenic in 3 tumors, isoechogenic in 2 and slightly hyperechogenic in 3. Heteroechogenicity was shown in 3 cases and homoechogenicity in 5. None of the margins was sharp. CT scan showed 4 isodense, 1 hypodense and 1 slightly hyperdense masses. After contrast enhancement, increased attenuation to different degrees but still lower than the normal renal parenchyma was observed. The sensitivity of IVP was low because of the small size of the tumor, peripheral location and rare involvement of the renal calyces and pelvis. MRI is helpful in differential diagnosis of hilar adenopathy and small vessels for tumor staging. Yet, the value of MRI for renal masses should be further studied. On cost benefit, sonography and CT are more practical in the diagnosis of renal carcinoma.

[Prognostic factors of primary non-Hodgkin's lymphoma of the stomach--clinicopathologic study of 21 cases].

Clinical, X-ray and pathological materials of 21 cases of primary non-Hodgkin's lymphoma (NHL) of the stomach were reviewed. According to NHL classification--working formulation for clinical usage sponsored by NCI, USA, there were 13 (61.9%) intermediate grade, 6 (28.6%) high grade and 2 (9.5%) low grade malignant lymphomas. By X-ray manifestations, they were classified into 5 types: multinodular type (4 cases), infiltrative (4), ulcerative (3), giant rugal (1) and mixed (9). Clinically, 10 lesions were stage I, 4 stage II, 1 stage III and 4 stage IV, 2 unclassified. The median survivals were 101 months for stage I, 15 months for stage II and less than 6 months for stages III and IV. The 5 and 10 year survival rates were 70% and 40% in stage I patients. The prognosis is related to clinical stage, depth of the tumor infiltration and histology type but not related to the tumor size or X-ray manifestations.

[X-ray features of primary non-squamous cell carcinoma and other malignant neoplasms in the trachea and main bronchi--analysis of 23 cases].

X-ray features of 23 cases (25 foci) of non-squamous cell carcinoma and other malignant neoplasms in the trachea and main bronchi were reviewed. They were 15 (17 foci) adenoid cystic carcinomas, 3 carcinoids, 2 mucoepidermoid carcinomas, 1 well-differentiated adenocarcinoma, leiomyosarcoma and extramedullary plasmocytoma each. The symptoms were non-specific leading to a delayed diagnosis more than one year in 52% of cases. The X-ray findings were classified into 3 types: intraluminal polypoid (11 tumors), sessile mass without and with extraluminal invasion (3 and 11 tumors). These tumors were prone to extraluminal invasion and can be demonstrated by X-ray. The air lumen involved appeared as localized expansion in 3 adenoid cystic carcinomas, 1 of which was proved by operation. Routine chest films were of limited value with 30% false negative chest film, 26% mediastinum mass and 30% obstructive pneumonitis/atelectasis. Lesions of posterior tracheal wall and carina were better revealed by lateral tomography. Of the 10 cases with lateral tomography, images were superior to those of AP tomography in 5. Three cases had CT scan, by which intra-tracheal/bronchial lesions, invasions of mediastinum and regional lymph nodes were shown. CT scan is more accurate compared with the other imaging modalities in visualizing these lesions and more helpful in selecting treatment.

[Malignant fibrous histiocytoma (MFH) of the maxilla--an analysis of 7 cases].

Malignant fibrous histiocytoma is a rare tumor of the soft tissue or bone, especially if located in the head and neck. Up to the present, only 7 cases have been reported (6 in the mandible and only one in the maxilla). From Mar. 1958 to Mar. 1985, 7 patients with maxillary MFH, proved by pathology, were treated in our hospital. The X-ray manifestations were studied and analysed. The main X-ray characteristics are as follows: A large soft tissue mass in the maxilla, extending into the infratemporal fossa, pterygopalatina fossa, orbit, ethmoid sinuses, nasal cavity and cheek. Marked and extensive destruction of the maxilla with some amorphous reactive ossification. The bone appears like melting ice or ground glass. The value of computerized tomography in differential diagnosis and clinical staging of the maxillary malignant fibrous histiocytoma are discussed.

Cranial computed tomography of breast carcinoma.

Cranial computed tomographic (CCT) manifestations of brain metastases were reviewed in 304 patients with carcinoma of the breast. Metastases were demonstrated in 103 patients; 13 had other significant abnormalities. Single lesions were found in 55 (54%) patients (cerebral hemisphere, 48; cerebellar hemisphere, 4; pineal, 1; pituitary and optic chiasm, 1; vermis, 1) and multiple metastases were found in 48 patients. Continuity with bony metastases was seen in 4 patients. Prior to contrast infusion, 13 (12.5%) patients with metastases had a normal CCT, and 13 (12.5%) showed lesions of increased attenuation. All metastases demonstrated enhancement following contrast infusion; 74 (72%) had a zone of surrounding "edema." Histopathologic-CCT correlation was available in 54 patients. Precontrast, increased attenuation of metastases was associated with hemorrhage or calcification, or both. Calcification accompanied by necrosis was the result of previous treatment. Following therapy, 3 patients demonstrated lesions that were impossible to differentiate from metastases by CCT, but proved histologically to have no viable tumor cells present.