The tumor immune microenvironment of nasopharyngeal carcinoma after gemcitabine plus cisplatin treatment.

Gemcitabine plus cisplatin (GP) chemotherapy is the standard of care for nasopharyngeal carcinoma (NPC). However, the mechanisms underpinning its clinical activity are unclear. Here, using single-cell RNA sequencing and T cell and B cell receptor sequencing of matched, treatment-naive and post-GP chemotherapy NPC samples (n = 15 pairs), we show that GP chemotherapy activated an innate-like B cell (ILB)-dominant antitumor immune response. DNA fragments induced by chemotherapy activated the STING type-I-interferon-dependent pathway to increase major histocompatibility complex class I expression in cancer cells, and simultaneously induced ILB via Toll-like receptor 9 signaling. ILB further expanded follicular helper and helper type 1 T cells via the ICOSL-ICOS axis and subsequently enhanced cytotoxic T cells in tertiary lymphoid organ-like structures after chemotherapy that were deficient for germinal centers. ILB frequency was positively associated with overall and disease-free survival in a phase 3 trial of patients with NPC receiving GP chemotherapy ( NCT01872962 , n = 139). It also served as a predictor for favorable outcomes in patients with NPC treated with GP and immunotherapy combined treatment (n = 380). Collectively, our study provides a high-resolution map of the tumor immune microenvironment after GP chemotherapy and uncovers a role for B cell-centered antitumor immunity. We also identify and validate ILB as a potential biomarker for GP-based treatment in NPC, which could improve patient management.

Fluorescence imaging and Raman spectroscopy applied for the accurate diagnosis of breast cancer with deep learning algorithms.

Deep learning is usually combined with a single detection technique in the field of disease diagnosis. This study focused on simultaneously combining deep learning with multiple detection technologies, fluorescence imaging and Raman spectroscopy, for breast cancer diagnosis. A number of fluorescence images and Raman spectra were collected from breast tissue sections of 14 patients. Pseudo-color enhancement algorithm and a convolutional neural network were applied to the fluorescence image processing, so that the discriminant accuracy of test sets, 88.61%, was obtained. Two different BP-neural networks were applied to the Raman spectra that mainly comprised collagen and lipid, so that the discriminant accuracy of 95.33% and 98.67% of test sets were gotten, respectively. Then the discriminant results of fluorescence images and Raman spectra were counted and arranged into a characteristic variable matrix to predict the breast tissue samples with partial least squares (PLS) algorithm. As a result, the predictions of all samples are correct, with minor error of predictive value. This study proves that deep learning algorithms can be applied into multiple diagnostic optics/spectroscopy techniques simultaneously to improve the accuracy in disease diagnosis.

Single-cell transcriptomics reveals regulators underlying immune cell diversity and immune subtypes associated with prognosis in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is an aggressive malignancy with extremely skewed ethnic and geographic distributions. Increasing evidence indicates that targeting the tumor microenvironment (TME) represents a promising therapeutic approach in NPC, highlighting an urgent need to deepen the understanding of the complex NPC TME. Here, we generated single-cell transcriptome profiles for 7581 malignant cells and 40,285 immune cells from fifteen primary NPC tumors and one normal sample. We revealed malignant signatures capturing intratumoral transcriptional heterogeneity and predicting aggressiveness of malignant cells. Diverse immune cell subtypes were identified, including novel subtypes such as CLEC9A+ dendritic cells (DCs). We further revealed transcriptional regulators underlying immune cell diversity, and cell-cell interaction analyses highlighted promising immunotherapeutic targets in NPC. Moreover, we established the immune subtype-specific signatures, and demonstrated that the signatures of macrophages, plasmacytoid dendritic cells (pDCs), CLEC9A+ DCs, natural killer (NK) cells, and plasma cells were significantly associated with improved survival outcomes in NPC. Taken together, our findings represent a unique resource providing in-depth insights into the cellular heterogeneity of NPC TME and highlight potential biomarkers for anticancer treatment and risk stratification, laying a new foundation for precision therapies in NPC.

CT and MRI findings in relapsing primary malignant melanoma of the lacrimal sac: a case report and brief literature review.

BACKGROUND: Primary lacrimal sac melanoma is an extremely rare condition with fewer than 50 cases reported so far. Clinically, its symptoms resemble those of dacryocystitis, leading to frequent misdiagnosis. During diagnosis, imaging examination is often performed to differentiate tumors from inflammation. In this report, we present a case of primary lacrimal sac melanoma and summarize the CT and MRI characteristics of lacrimal sac melanoma. CASE PRESENTATION: We report a 50-year-old female patient who had undergone a dacryocystectomy for the left lacrimal sac mass. Postoperative pathological examination confirmed the presence of primary malignant melanoma. Three months later, a lump in the lacrimal sac area was found. The patient underwent CT and MR examinations. CT scan demonstrated a partially well-defined soft mass in the fossa of left lacrimal sac extending into the nasolacrimal duct and anterior ethmoid sinus. MRI revealed an intermediate signal intensity on T1 and iso-or hyper-signal on T2 weighted images. Histopathological examination on biopsy confirmed recurrence of primary lacrimal sac melanoma. DISCUSSION AND CONCLUSIONS: None has described the CT and MR findings of primary lacrimal sac melanoma so far. Typically, MR images show hyperintensity signal on T1-weighted images and hypointense signal on T2-weighted images owing to the paramagnetic properties of melanin. In contrast to previous reports and the present case, most cases do not present these typical signals. Thus, reporting such radiological findings is important to create awareness on variant images of primary lacrimal sac melanoma. This will reduce misdiagnosis and mistreatment.

Prediction and prophylaxis of hepatocellular carcinoma occurrence and postoperative recurrence in chronic hepatitis B virus-infected subjects.

Hepatocellular carcinoma (HCC) is one of the most common and highly fatal malignancies worldwide. Chronic infection with hepatitis B virus (HBV) is a major cause of HCC. High HBV replication rate and related non-resolving inflammation are the major risk factors of HCC occurrence and postoperative recurrence. Early prophylactic options are effective in reducing HCC occurrence and improving survival. Therefore, it is important to identify HBV-infected patients who are at a higher risk of developing HCC and HBV-HCC patients who are more likely to relapse after surgery, thus providing them with more precise prophylactic strategies. Several prediction models of HCC occurrence have been constructed, with satisfactory predictive accuracy and discriminatory ability. However, there is a lack of consensus for their clinical implementation. Several staging systems have been proposed for HCC prognosis. However, the accuracy of these staging systems based on demographic characteristics and clinical measurements needs to be further improved, possibly by systematically incorporating viral and inflammatory factors. Since antiviral treatments are effective in promoting liver function reserve, reducing HCC occurrence and prolonging postoperative survival in some HBV-infected subjects, it is very important to identify subgroups of HBV-infected patients who would most benefit from antiviral treatment.

Systemic genome screening identifies the outcome associated focal loss of long noncoding RNA PRAL in hepatocellular carcinoma.

UNLABELLED: Systemic analyses using large-scale genomic profiles have successfully identified cancer-driving somatic copy number variations (SCNVs) loci. However, functions of vast focal SCNVs in "protein-coding gene desert" regions are largely unknown. The integrative analysis of long noncoding RNA (lncRNA) expression profiles with SCNVs in hepatocellular carcinoma (HCC) led us to identify the recurrent deletion of lncRNA-PRAL (p53 regulation-associated lncRNA) on chromosome 17p13.1, whose genomic alterations were significantly associated with reduced survival of HCC patients. We found that lncRNA-PRAL could inhibit HCC growth and induce apoptosis in vivo and in vitro through p53. Subsequent investigations indicated that the three stem-loop motifs at the 5' end of lncRNA-PRAL facilitated the combination of HSP90 and p53 and thus competitively inhibited MDM2-dependent p53 ubiquitination, resulting in enhanced p53 stability. Additionally, in vivo lncRNA-PRAL delivery efficiently reduced intrinsic tumors, indicating its potential therapeutic application. CONCLUSIONS: lncRNA-PRAL, one of the key cancer-driving SCNVs, is a crucial stimulus for HCC growth and may serve as a potential target for antitumor therapy.

Huachansu injection inhibits metastasis of pancreatic cancer in mice model of human tumor xenograft.

BACKGROUND: Huachansu injection (HCS) is a water-soluble preparation made from Bufo gargarizans's skin, which has been widely used in clinics for tumor therapy in China. Though the anti-cancer activity of HCS has been verified through studies in vitro and in vivo, there is little research about its potential anti-metastasis effect. The primary objective of this study was to assess the effects of HCS on both the invasion of pancreatic cancer cells in vitro and on the progression of liver metastasis in vivo in this study. METHODS: HCS anti-metastasis potential was accessed using both assay of Cell viability and invasion in vitro, and then further Establishing xenograft model in nude mice. In the cell-based assay, mRNA and protein expression of MMP-2, MMP-9 and VEGF was detected by semi-quantitative RT-PCR and western blotting. In animal experiment, liver metastasis nodules and change of liver-body ratio was observed. Meanwhile, correlation of the CA19-9 and CEA content in serum with the progression of liver metastasis was analyzed. RESULT: We observed that HCS prevented the invasion of cancer cells, with inhibiting the expressions of MMP-2 and MMP-9, and reduced not only the number of metastasis nodules but the ratio of liver-body weight as well. Furthermore, HCS decreased the expression of MMP-2, MMP-9 and VEGF in liver metastasis, while also reducing CA19-9 contents in serum. In addition, correlation analysis indicated that the level of CA19-9 in serum was closely related to the number of liver metastasis nodules. CONCLUSION: Our experimental results suggest that HCS has some anti-metastasis potential to suppress the growth of liver metastasis by decreasing the expression of MMP-2 and MMP-9 as well as VEGF.

[Applications and progress of Fourier transform infrared spectroscopic microimaging in bone disease research].

Fourier transform infrared spectroscopy (FTIRS) and microimaging technique have been integrated together to evolve into Fourier transform infrared spectroscopic imaging (FTIRI) system. This system can provide not only the morphological information of the sample by visible image and FTIR image, but also the abundant information on the spectral, component and structure of specimen by FTIRS, especially of the heterogeneous solid mixture. The richer and more visualized information obtained by FTIRI greatly raised the research efficiency and usability of the spectral technique in biomedicine, pharmacology, forensic medicine, material science and chemistry, etc. The present paper depicts FTIRI development process, system structure, imaging principle and mode selection; and then introduces that FTIRI opened a new area of investigation for biomedicine, namely, research on bone disease by FTIRI. Then the paper illustrates the related research findings and progress in FTIRI use for osteopetrosis, osteogenesis imperfecta, osteoporosis and osteomalacia, as well as a couple of limitations. The prospective study for FTIRI in biomedical research field is also addressed.

[Apoptosis of A549 cells induced by cloned Noxa gene].

OBJECTIVE: To clone the Noxa gene and to observe the apoptosis of A549 cells transfected with the recombinant plasmid of pcDNA-Noxa. METHODS: The Noxa gene was obtained by PCR, and was cloned into pcDNA3. 1(-). A549 cells were transfected with the recombinant plasmid of pcDNA-Noxa. Western blot analysis was performed to determine the overexpression of Noxa. A549 cells were stained with Hoechst 33258 to observe the apoptosis. RESULTS: The recombinant plasmid of pcDNA-Noxa was successfully constructed evidenced by endonuclease digestion and sequence analysis. The overexpression of Noxa was identified using Western blot analysis. The recombinant plasmid of pcDNA-Noxa induced apoptosis of A549 cells. CONCLUSION: Nora has exhibited potential pro-apoptotic activity against A549 cells. This study is a foundation for further research into pro-apoptotic activity of Noxa gene.

[Association of genetic polymorphisms of key molecules in JAK/STAT signaling pathway with susceptibility of hepatocellular carcinoma].

OBJECTIVE: To elucidate the association of genetic polymorphisms of key molecules in JAK/STAT signaling pathway with susceptibility of hepatocellular carcinoma (HCC). METHODS: A total of 367 HCC patients and 367 healthy controls were recruited in this sex- and age-matched case-control study. Genetic polymorphisms of IL-6 (rs1800796, -572C > G), STAT3 (rs744166, +26312T > C; rs3816769, +42240T > C; rs6503695, +40980T > C), EGFR (rs11543848, +142530A > G), and mTOR (rs7211818, +170278A > G; rs9674559, +196983A > G; rs11653499, +65678G > A) were genotyped using a mass spectrometry method. Odds ratio (OR) and 95% confidence interval (CI) were calculated. RESULTS: Genotype frequency of the 8 polymorphisms of IL-6, STAT3, EGFR, and mTOR were not significantly different between the patients with HCC and the controls. When stratified by sex, the female subjects who carried STAT3 +26312CC, +42240CC, or +40980CC had a decreased risk of HCC when compared to those who carried TT allele (OR = 0.192, 95%CI: 0.047 - 0.784; OR = 0.180, 95%CI: 0.045 - 0.725; OR = 0.198, 95%CI: 0.049 - 0.806, respectively). When compared with AA genotype on the site of EGFR +142530, the (AG + GG) genotype reduced the risk of HCC in women (OR = 0.422, 95%CI: 0.179 - 0.994). CONCLUSION: The polymorphisms of IL-6 (rs1800796) and mTOR (rs7211818, rs9674559, and rs11653499) were not associated with the HCC susceptibility. Those carrying CC allele in three loci (rs744166, rs3816769, and rs6503695) of STAT3 and (AG + GG) in rs11543848 of EGFR had a decreased risk of HCC in women. However, these results need to be validated using larger sample size.

Qingyihuaji formula inhibits progress of liver metastases from advanced pancreatic cancer xenograft by targeting to decrease expression of Cyr61 and VEGF.

OBJECTIVE: To observe the effects of Qingyihuaji formula (QYHJ) on the progression of liver metastases from human pancreatic cancer and to detect the expression changes of some biological factors associated with angiogenesis and metastasis during the development of advanced pancreatic cancer. METHODS: Nude mice were inoculated intrasplenically with human pancreatic cancer cell line SW1990 and then randomly assigned into 4 groups: a control group and groups QYHJ-A, QYHJ-B, and QYHJ-C. Following this, the mice were treated with or without QYHJ formula for 4 weeks and were sacrificed at the end of the sixth week. The changes in body weight were observed, followed by the livers being excised and weighed. Then, both the numbers and the volume of metastatic nodules per liver were evaluated. Subsequently, the expressions of MMPs, VEGF, and Cyr61 in the tissue of liver metastases were detected by reverse transcription polymerase chain reaction, immunohistochemistry, or Western blot. Finally, the correlation was evaluated between the expressions of the factors associated with metastasis and the growth of liver metastasis. RESULTS: Liver metastases were identified in 11 of 15 mice (73%) in the control group, 9 of 15 mice (60%) in group QYHJ-A, 6 of 14 mice (43%) in group QYHJ-B, and 8 of 14 mice (57%) in group QYHJ-C both the number and the volume of metastatic nodules per liver same as the ratio of liver-body weight in QYHJ groups were significantly less than the controlled group (P < 0.05). The expressions of Cyr61, MMP-2, and VEGF at the levels of mRNA and protein were decreased in the QYHJ groups when compared with the control, as confirmed by immunohistochemistry detection (P < .05). However, no significant difference was observed in the mRNA expression of MMP-1 and MMP-9 between the QYHJ groups and the control group (P > .05). Regression analysis indicated that QYHJ possessed an evident inhibition against the progression of liver metastasis by downregulating the expression of VEGF and Cyr61 rather than MMP-2. CONCLUSIONS: The QYHJ formula exerted an inhibitory effect on the growth of liver metastasis from pancreatic cancer, perhaps by targeting VEGF and Cyr61 to some extent.

Factors predicting occurrence and prognosis of hepatitis-B-virus-related hepatocellular carcinoma.

Primary liver cancer is an important cause of cancer death, and hepatocellular carcinoma (HCC) accounts for 70%-85% of total liver cancer worldwide. Chronic hepatitis B virus (HBV) infection contributes to > 75% of HCC cases. High serum viral load is the most reliable indicator of viral replication in predicting development of HCC. HBV genotype C is closely associated with HCC in cirrhotic patients aged > 50 years, whereas genotype B is associated with development of HCC in non-cirrhotic young patients and postoperative relapse of HCC. Different HBV subgenotypes have distinct patterns of mutations, which are clearly associated with increased risk of HCC. Mutations accumulate during chronic HBV infection and predict occurrence of HCC. Chronic inflammation leads to increased frequency of viral mutation via cellular cytidine deaminase induction. Mutations are negatively selected by host immunity, whereas some immuno-escaped HBV mutants are active in hepatocarcinogenesis. Inflammatory pathways contribute to the inflammation-necrosis-regeneration process, ultimately HCC. Their hallmark molecules can predict malignancy in HBV-infected subjects. Continuing inflammation is involved in hepatocarcinogenesis and closely related to recurrence and metastasis. HBV load, genotype C, viral mutations and expression of inflammatory molecules in HBV-related HCC tissues are significantly associated with poor prognosis. Imbalance between intratumoral CD8(+) T cells and regulatory T cells or Th1 and Th2 cytokines in peritumoral tissues can predict prognosis of HBV-related HCC. These factors are important for developing active prevention and surveillance of HBV-infected subjects who are more likely to develop HCC, or for tailoring suitable treatment to improve survival or postpone postoperative recurrence of HCC.

Association of novel mutations and haplotypes in the preS region of hepatitis B virus with hepatocellular carcinoma.

The association of viral mutations and haplotypic carriages with mutations in the preS region of hepatitis B virus (HBV) genotypes B and C with hepatocellular carcinoma (HCC) is of great significance for the prediction of this malignancy, but it remains obscure. We analyzed the preS sequences of HBV genotypes B and C from 1172 HBV-infected subjects including 231 patients with HCC. As compared with the HBV-infected subjects without HCC, C2875T, G2946C, A3054C, C3060A, T3066C, C3116T, A3120C, G3191A, A1C, C7A, C10A, A31C, C76T, G105C, and G147C in both genotypes were significantly associated with increased risks of HCC. C2875A, G2950A, G2951A, A3054T, C3060T, T3066A, T3069G, A3120T, and G3191C were significantly associated with increased risks of HCC in genotype C, whereas these mutations were inversely associated with HCC in genotype B. Multivariate regression analyses showed that C76A/T was a novel factor independently associated with an increased risk of HCC, as compared with those without HCC. The frequencies of haplotypes 2964A-3116T-preS2 start codon wild-type-7C, 2964C-3116T-7A-76C, and 2964A-3116T-7C-76A/T were significantly higher in the patients with HCC (P<0.001), whereas a haplotypic carriage with a single mutation and another three wildtypes were inversely associated with HCC. Conclusively, the association of HBV mutations in the preS region with HCC depends on HBV genotype and haplotypic carriage with two or more mutations that are each associated with an increased risk of HCC independently.

Diagnostic value of whole body diffusion weighted imaging for screening primary tumors of patients with metastases.

OBJECTIVE: To evaluate the values of whole body diffusion weighted imaging (DWI) in screening primary unknown tumor in patients with metastases. METHODS: Totally, 34 patients with metastases of primary unknown tumors were scanned with whole body DWI, and conventional magnetic resonance (MR) imaging was performed if suspected lesions were detected. All the metastases including 27 cases of osseous metastases, 2 brain metastases, 2 liver metastases, 1 pulmonary multiple metastasis, 1 neck metastasis and 1 malignant ascites, were diagnosed by computed tomography, single photon emission computed tomography, or MR imaging. For the proven primary tumors diagnosed by biopsy or pathology of surgical specimens, apparent diffusion coefficient (ADC) values of the primary and metastatic lesions were measured respectively. The sensitivity and specificity of this technique for screening primary tumors were evaluated. RESULTS: We found 24 cases with suspected primary lesions, in which 23 lesions were proved to be primary tumors, and 1 was proved to be benign lesion. And no definite primary lesion was found in 10 cases on whole body DWI, but in which 1 case was diagnosed with primary tumor by biopsy later, and the other 9 cases remained unknown within follow-up of over half a year. The difference was not significant in ADC values between primary and metastatic lesions (P>0.05). The sensitivity and specificity of whole body DWI for searching primary tumors was 95.8% and 90.0%, respectively. CONCLUSION: Combined with conventional MR scanning, whole body DWI can help to search primary lesions of patients with metastases.