Interferon-γ induces tumor resistance to anti-PD-1 immunotherapy by promoting YAP phase separation.

Interferon-γ (IFN-γ)-mediated adaptive resistance is one major barrier to improving immunotherapy in solid tumors. However, the mechanisms are not completely understood. Here, we report that IFN-γ promotes nuclear translocation and phase separation of YAP after anti-PD-1 therapy in tumor cells. Hydrophobic interactions of the YAP coiled-coil domain mediate droplet initiation, and weak interactions of the intrinsically disordered region in the C terminus promote droplet formation. YAP partitions with the transcription factor TEAD4, the histone acetyltransferase EP300, and Mediator1 and forms transcriptional hubs for maximizing target gene transcriptions, independent of the canonical STAT1-IRF1 transcription program. Disruption of YAP phase separation reduced tumor growth, enhanced immune response, and sensitized tumor cells to anti-PD-1 therapy. YAP activity is negatively correlated with patient outcome. Our study indicates that YAP mediates the IFN-γ pro-tumor effect through its nuclear phase separation and suggests that YAP can be used as a predictive biomarker and target of anti-PD-1 combination therapy.

Missense variants in CYP4B1 associated with increased risk of lung cancer among Chinese Han population.

INTRODUCTION: Understanding the etiology and risk factors of lung cancer (LC) is the key to developing scientific and effective prevention and control strategies for LC. CYP4B1 genetic polymorphism has been reported to be associated with susceptibility to various diseases. We aimed to explore the association between CYP4B1 genetic variants and LC susceptibility. METHODS: One thousand three hundred thirty-nine participants were recruited to perform an association analysis through SNPStats online software. Statistical analysis of this study was mainly completed by SPSS 22.0 software. False-positive report probability analysis (FPRP) to detect whether the positive findings were noteworthy. Finally, the interaction of SNP-SNP in LC risk was evaluated by multi-factor dimensionality reduction. RESULTS: We found evidence that missense variants in CYP4B1 (rs2297810, rs4646491, and rs2297809) are associated with LC susceptibility. In particular, genotype GA of CYP4B1-rs2297810 was significantly associated with an increased risk of LC in both overall and stratified analyses (genotype GA: OR (95% CI) = 1.35 (1.08-1.69), p = 0.010). CYP4B1-rs4646491 (overdominant: OR (95% CI) = 1.30 (1.04-1.62), p = 0.023) and CYP4B1-rs2297809 (genotype CT: OR (95% CI) = 1.26 (1.01-1.59), p = 0.046) are also associated with an increased risk of LC. FPRP analysis showed that all positive results in this study are noteworthy findings CONCLUSION: Three missense variants in CYP4B1 (rs2297810, rs4646491, and rs2297809) are associated with increasing risk of LC.

Characteristics of preplan-based three-dimensional individual template-guided brachytherapy compared to freehand implantation.

Image-guided adaptive intracavitary/interstitial brachytherapy (IC/IS IGABT) has exhibited superior dosimetry advantage and local control for locally advanced cervical cancer (LACC). Our group designed a type of cylindrical three-dimensional (3D) printed vaginal template combining an intracavitary applicator with straight and oblique interstitial needles according to the preplan on computed tomography images. This work aimed to research the consistency of the preplan with the treatment plan at every fraction to verify the practical guiding significance of the preplan. We also investigated the difference between 3D-printed template-guided implantation compared with freehand implantation for LACC. Twenty-six patients were treated with 3D-printed individual templates (3D template group), and 20 patients were treated by using freehand insertion (freehand group). Patients in the 3D template group would take a preplan one week before treatment to design and print the individual template, while the freehand group did not. All patients accepted volumetric rotational intensity-modulated radiotherapy at a dose of 49.4 Gy in 26 fractions and subsequent brachytherapy at a dose of 26 Gy in four fractions. All analyses were performed by utilizing SPSS 26. The insertion depth was decreased in fractions 1 and 4 compared with the preplan. None of the dose volume histogram parameters of fractions 1-3, nor the D2cc of bladder and bowel at fraction 4 were barely changed compared with the preplan. The D90 and D98 of the high-risk clinical target volume in the 3D template group were statistically higher than those in the freehand group (p < 0.01). The D2cc of the rectum, bladder, bowel, and sigmoid in the 3D template group were all lower than those in the freehand group (p < 0.01). The preplan in this research is consistent with treatment plans, which is important to ensure the feasibility of applying a 3D-printed template in brachytherapy. The 3D-printed individual guidance template was an effective method in brachytherapy for locally advanced cervical cancer.

Effect of genetic distances of different genotypes of maize on the authenticity of single seeds detected by NIR spectroscopy.

Introduction: NIR spectroscopy combined with chemometric algorithms has been widely used for seed authenticity detection. However, the study of seed genetic distance, an internal feature that affects the discriminative performance of classification models, has rarely been reported. Methods: Therefore, maize seed samples of different genotypes were selected to investigate the effect of genetic distance on the authenticity of single seeds detected by NIR spectroscopy. Firstly, the Support vector machine (SVM) model was established using spectral information combined with a preprocessing algorithm, and then the DNA of the samples was extracted to study the correlation between genetic and relative spectral distances, the model identification performance, and finally to compare the similarities and differences between the results of genetic clustering and relative spectral clustering. Results: The results were as follows: the average accuracy of the models was 93.6% (inbred lines) and 93.7% (hybrids), respectively; Genetic distance and correlation spectral distance exhibited positive correlation significantly (inbred lines: r=0.177, p<0.05; hybrids: r=0.238, p<0.05), likewise genetic distance and model accuracy also showed positive correlation (inbred lines: r=0.611, p<0.01; hybrids: r=0.6158, p<0.01); Genetic clustering and spectral clustering results were essentially uniform for 94.3% (inbred lines) and 93.9% (hybrids), respectively. Discussion: This study reveals the relationship between the genetic and relative spectral distances of seeds and the accuracy of the model, which provides theoretical basis for the establishment of the standardized system for detecting the authenticity of seeds by NIR spectroscopic techniques.

[Effects of berberine on LPS /NF-κB and MAPK signaling pathways in PCOS model rats].

Objective: To investigate the effects of berberine on glucose and lipid metabolism, sex hormone binding protein, adiponectin (LPS), NF-κB and MAPK signaling pathways in polycystic ovary syndrome (PCOS) model rats. Methods: Female SD rats were randomly divided into control group, PCOS model group, berberine (0.216 g/kg) group, metformin (0.135 g/kg) group and Dyne-35 (0.18 mg/kg) group, 10 rats in each group. The rats in PCOS model group were treated with letrozole 1 mg.kg-1 by ig for 3 weeks. After 28 days of drug intervention, the body constitution, ovarian and uterine indexes of the rats were detected, and the changes in the number of ovarian follicles were observed by HE staining. The levels of serum sex hormone, glucose and insulin, triglyceride and cholesterol, sex hormone-binding protein and adiponectin were determined by ELISA, and the protein expressions of p38-MAPK, C-Jun and NF-κB in ovarian tissues were determined by Western blot. Results: Compared with control group, body weight of model group was increased (P＜0.05), and uterine index was decreased (P＜0.05); The number of follicles was increased (P＜0.05). Serum levels of luteinizing hormone (LH), testosterone (T) and LH/FSH ratio were increased (P＜0.05), follicular estrogen (FSH) level was decreased (P＜0.05), total cholesterol (TC), triglyceride (TG), fasting insulin and insulin index (HOMA) were increased (P＜ 0.05). The content of sex hormone binding protein (SHBG) was decreased and the content of adiponectin (LPS) was increased (P＜0.05). The protein expressions of p38-MAPK, c-Jun and NF-κB in ovarian tissue were up-regulated (P＜0.05). Compared with model group, in berberine group, the uterine index and the number of secondary follicles were increased(P＜0.05), the serum levels of luteinizing hormone (LH) , testosterone (T) and the ratio of LH/FSH were decreased (P＜0.05), and the protein expression levels of p38-MAPK and NF-κB in ovarian tissue were down-regulated (P＜0.05), which were similar to those of Dyne-35 group. Berberine significantly decreased serum triglyceride (TG), insulin level and insulin index (P＜0.05), increased serum SHBG level and decreased serum LPS level (P＜0.05), which were similar to those of metformin. Conclusion: Berberine can regulate sex hormone disorder and insulin resistance (IR) in PCOS rats by down-regulating the expressions of p38-MAPK and NF-κB protein in ovarian tissues and decreasing the serum content of LPS.

Massive prolapsed cervical lumps in cervical cancer treated with three-dimensional implant brachytherapy: a case report.

Background: Cervical cancer is one of the most common gynecological malignancies in developing countries. But the cervical cancer patients with tumor prolapse are very rare. The treatment principle of cervical cancer has been written into the guide, while the management of cervical lumps prolapse associated with cervical cancer is not standardized. Every doctor has different opinions on treatment strategies. Herein, we describe the three-dimensional brachytherapy treatment for massive prolapsed cervical lumps. Case Description: A 48-year-old woman diagnosed with cervical cancer developed a huge cervical mass prolapsed after defecation on the second day of chemotherapy. The mass surface was continuously bleeding and unable to return to the vagina. Therefore, uterine artery embolization interventional hemostasis was performed and then three-dimensional brachytherapy treatment was applied. The mass was necrotic and shedding and then retracted into the vagina on the 7th day after implantation treatment. Finally, the patient successfully received radical radiotherapy [pelvic and abdominal cavity external beam radiotherapy-PCTV 50.4 Gy/28 F, pelvic metastatic lymph nodes PGTVn 61.0 Gy/28 F, plus vaginal three-dimensional brachytherapy-HRCTV (D90) 27.25 Gy/4 F]. Conclusions: If cervical cancer combined with tumor prolapse is inoperable, three-dimensional implants brachytherapy seems to be an adequate therapeutic option.

Plasma levels of methylated septin 9 are capable of detecting hepatocellular carcinoma and hepatic cirrhosis.

Hepatocellular carcinoma (HCC) was the third most common cause of cancer­associated mortality in China in 2015. Early detection of HCC and hepatic cirrhosis (HC) can serve a crucial role in the prevention and therapeutic intervention of these diseases. Current early detection methods rely on less sensitive imaging modalities compared with the pathological examination. In the present study, a total of 64 patients with HCC, 44 patients with HC and 298 individuals with no evidence of disease (NED) were recruited, and the ability of methylated septin 9 (mSEPT9) in diagnosing HCC and HC was investigated. The overall detection sensitivity of mSEPT9 for HCC and HC was 76.7 and 34.1%, respectively, with a 95.9% specificity (HCC vs. NED). The sensitivity of mSEPT9 for HCC was significantly higher than that of α­fetoprotein (AFP; χ2 test; 56.7%; P<0.05). The areas under the curve from the receiver operating characteristic curves of mSEPT9 for detection of HCC vs. NED, HC vs. NED and HCC vs. HC were 0.85, 0.77 and 0.66, respectively, while those of AFP for the same groups were 0.80, 0.55 and 0.77, respectively. Although both markers exhibited stage­dependent sensitivity in HCC, mSEPT9 was demonstrated to be more sensitive than AFP. The net reclassification index of mSPET9 for HCC detection was 0.212 compared with AFP, suggesting an improved diagnostic performance of mSEPT9 compared with AFP. In addition, Kaplan­Meier survival analysis revealed that mSEPT9 is able to predict the long­term survival of patients with HCC. Further analysis suggested that patients >50 years of age exhibited higher sensitivity compared with those <50 years old in mSEPT9, but not in AFP. No significant difference in sensitivity was observed between compensated and decompensated patients with HC, and in patients with HC with a history of hepatitis B or C virus infection. No difference was observed between male and female subjects in the HC and HCC groups for mSEPT9 and AFP. In conclusion, mSEPT9 may detect HCC with an overall improved sensitivity compared with AFP and may help in predicting the long­term survival of patients with HCC. The present clinical study was retrospectively registered to the Chinese Clinical Trial Registry on April 4, 2020 (http://www.chictr.org.cn/enIndex.aspx; registration no. ChiCTR2000031547).

A comparative dosimetric study of cervical cancer patients with para-aortic lymph node metastasis treated with volumetric modulated arc therapy vs. 9-field intensity-modulated radiation therapy.

BACKGROUND: To compare the dosimetric characteristics between volumetric modulated arc therapy (VMAT) and 9-field intensity-modulated radiation therapy (9F-IMRT) for cervical cancer patients with para-aortic lymph node (PALN) metastasis. METHODS: We selected 20 patients who had received extended-field radiotherapy for cervical cancer with PALN metastasis. IMRT and VMAT plans were compared in terms of target, organs at risk (OARs), homogeneity index (HI), conformity index (CI), the number of monitor units (MUs) and treatment time (s). RESULTS: The CI and HI of VMAT plans were superior to those of IMRT plans (P<0.05). As for OARs, the mean maximum doses (Dmean) to the kidneys in the VMAT plans were all lower than those in IMRT plans (P<0.001). V40, V50 of the rectum, and V40 of the bladder in VMAT plans involved fewer doses than IMRT plans (P<0.001). Compared with IMRT plans, VMAT reduced the average number of MUs by 51% and the average treatment time by 31%. CONCLUSIONS: Both VMAT and IMRT plans can satisfy clinical dosimetric demands and protect OARs. VMAT has the best performance on CI and HI and can better protect the OARs. VMAT plans have fewer MUs and improve treatment efficiency.

Wnt1 and Wnt2b immunodetection of the regenerating tail and comparative ultrastructure of tail spinal cord in the Scincella tsinlingensis / Inmunodetección Wnt1 y Wnt2b de la cola en regeneración y ultraestructura comparativa de la médula espinal de la cola en Scincella tsinlingensis

SUMMARY: The Wnt pathway is essential for the initiation of lizard tail regeneration. The regenerated lizard tails exhibit obvious morphological differences compared to the original ones. The expression of Wnt1 and Wnt2b proteins in the regenerating tail of Scincella tsinlingensis was detected by immunohistochemistry and then comparatively analyzed for ultrastructural changes in the original and regenerated spinal cord. The ependymal layer of the original spinal cord was pseudostratified with multiciliated cells and primary monociliated cells, while the cells of the ependymal layer of the regenerated spinal cord were organized in a monolayer with a few biciliated cells. Immunolocalization indicated that Wnt1 and Wnt2b were mainly distributed in the dermis near the original tail stump, spinal cord, and clot-positive migratory cells during Stage I, 0-1 days post-amputation (dpa). Wnt1 and Wnt2b were predominantly detected in the epaxial and hypaxial musculature near the original tail stump, wound epithelium, and spinal cord in the original tail during Stage II, 1-7 dpa. Mesenchymal cells and wound epithelium showed immunostaining during Stage III and IV, 7-15 dpa. The ependymal tubes contained these signaling proteins during Stage V and VI, 20- 30 dpa. Labeling was mainly observed in nearby regenerative blood vessels, ependymal cells, epaxial and hypaxial musculature in the apical epithelial layer (AEC) after 45-160 dpa. These findings indicated that Wnt1 and Wnt2b proteins presented primarily in regenerating epidermis and nerve tissues were a critical signal for tail regeneration in S. tsinlingensis.

RESUMEN: La vía Wnt es esencial para el inicio de la regeneración de la cola del lagarto. Las colas de lagarto regeneradas exhiben diferencias morfológicas obvias en comparación con las originales. La expresión de las proteínas Wnt1 y Wnt2b en la cola en regeneración de Scincella tsinlingensis se detectó mediante inmunohistoquímica y luego se analizaron comparativamente los cambios ultraestructurales en la médula espinal original y regenerada. La capa ependimaria de la médula espinal original se pseudoestratificó con células multiciliadas y células monociliadas primarias, mientras que las células de la capa ependimaria de la médula espinal regenerada se organizaron en monocapa con algunas células bicilicadas. La inmunolocalización indicó que Wnt1 y Wnt2b se distribuyeron principalmente en la dermis cerca del muñón de la cola original, la médula espinal y las células migratorias positivas en el coágulo durante la Etapa I, 0-1 días después de la amputación (dpa). Wnt1 y Wnt2b se detectaron predominantemente en la musculatura epaxial e hipaxial cerca del muñón de la cola original, el epitelio de la herida y la médula espinal en la cola original durante la Etapa II, 1-7 dpa. Las células mesenquimales y el epitelio de la herida mostraron inmunomarcaje durante la Etapa III y IV, 7- 15 dpa. Los tubos ependimarios contenían estas proteínas de señalización durante la Etapa V y VI, 20-30 dpa. El marcaje se observó principalmente en vasos sanguíneos regenerativos cercanos, células ependimarias, musculatura epaxial e hipaxial en la capa epitelial apical (AEC) después de 45-160 dpa. Estos hallazgos indicaron que las proteínas Wnt1 y Wnt2b están presentes principalmente en la epidermis en regeneración y en los tejidos nerviosos y eran una señal crítica para la regeneración de la cola en S. tsinlingensis.

[Eukaryotic expression and localization of human MTERF2 in cervical cancer Caski cells].

Objective To construct the recombinant eukaryotic expression vector of human mitochondrial transcription termination factor 2 (MTERF2) gene and determine the cellular localization by overexpressing MTERF2 in human Caski cervical cancer cells. Methods The coding sequence of MTERF2 was amplified by reverse transcription-PCR using the total RNA extracted from human cervical cancer Caski cells, and then was inserted into p3×FLAG-CMV-14 vector. After being verified by PCR and DNA sequencing, the positive recombinant plasmid was transiently transfected into Caski cells. Western blotting and immunofluorescence technique were performed to analyze the expression and distribution of human MTERF2 proteinat 24, 32 and 48 hours after transfection. Results Sequence analysis showed that the correct target gene (1158 bp) was inserted into the vector at the expected position. The target protein band was detected at Mr 44 000 as we had predicted in the transfected cells while not in the negative control group, which indicated MTERF2 expression vector could be successfully transfected and expressed in Caski cells. The p3×FLAG-MTERF2 protein was highly expressed and displayed a mitochondrial distribution at 24 hours post-transfection in Caski cells. Conclusion We successfully constructed the eukaryotic expression plasmid p3×FLAG-CMV-MTERF2 and expressed p3×FLAG tagged MTERF2 effectively in the mitochondria of Caski cells.

Expression of growth­regulated oncogene­1, hepatocyte growth factor, platelet­derived growth factor­AA and soluble E­selectin and their association with high­risk human papillomavirus infection in squamous cell carcinoma of the uterine cervix.

The aim of the present study was to evaluate the clinical significance and prognostic value of growth­regulated oncogene­1 (GRO­1), hepatocyte growth factor (HGF), platelet­derived growth factor­AA (PDGF­AA), soluble E­selectin (sE­selectin) and high­risk human papillomavirus (HPV; types 16, 18/45, 31 and 33/52/58/67) infection in cervical squamous cell carcinoma (CSCC). A total of 426 cases were enrolled in the present study, of which 292 cases were patients with CSCC, 43 were patients with cervical intraepithelial neoplasia (CIN) and 91 were healthy controls. Luminex xMAP technology was used to detect the serum levels of GRO­1, HGF, PDGF­AA and sE­selectin in all cases and two­channel fluorescence quantitative polymerase chain reaction was used to determine HPV DNA in cervical scrapings from CSCC and CIN patients. The results demonstrated that the serum levels of GRO­1, HGF and sE­selectin were significantly higher in patients with CSCC compared with patients with CIN and the healthy controls (P<0.0001). Compared with the CIN patients, the HPV positive rate in the CSCC patients significantly increased (P=0.013). The four factors were correlated with certain clinicopathological variables of CSCC patients to a certain degree (P<0.05) and the levels of HGF were closely associated with HPV infection (P=0.039). The receiver operating characteristic curves demonstrated that HGF obtained the highest diagnostic value compared with the other three factors. Multivariate Cox regression analysis demonstrated that the serum levels of HGF (P<0.0001), FIGO stage (P<0.0001) and pelvic lymph node metastasis (P=0.001) were independent prognostic factors in patients with CSCC, while high­risk HPV infection did not show any significance in this analysis. These results demonstrated that HGF may be a useful prognostic biomarker rather than high­risk HPV types in patients with CSCC.

Genome analysis and development of infectious cDNA clone of a virulence-attenuated strain of foot-and-mouth disease virus type Asia 1 from China.

The RNA genome sequence of the rabbit passage-attenuated strain of foot-and-mouth disease virus (FMDV) Asia 1, ZB/CHA/58(att), was determined to be 8165 nt in length excluding the poly(C) tract in the 5' UTR and the poly(A) tail at the 3' end. ZB/CHA/58(att) was most similar to the vaccine strain Asia 1/YNBS/58 in genome sequence and there were no deletions or insertions within the deduced polyprotein between ZB/CHA/58(att) and YNBS/58, but there were a total of 25 substitutions at the amino acid level and an extra 19-nt stretch in the 5' UTR was found in ZB/CHA/58(att). An infectious full-length cDNA clone of ZB/CHA/58(att) was developed. Infectious virus could be recovered in BHK-21 cells transfected with the synthetic viral RNA transcribed in vitro. The plaque morphology, growth kinetics and antigenic profile of the infectious clone-derived virus (termed tZB) were indistinguishable from those induced by the parental virus. Furthermore, the virulence properties of ZB/CHA/58(att) and tZB were found to be highly similar in the mouse model. The availability of genome sequence information and infectious cDNA clone of the FMDV ZB/CHA/58(att) lays a new ground for further investigation of FMDV virulence determinants and development of new potent vaccine to FMD.

[Effect of autologous bone marrow stem cell transplantation on renal function following renal ischemic-reperfusion in rabbits].

OBJECTIVE: To observe the effect of autologous bone marrow stem cell (BMSC) transplantation via the renal artery on renal function recovery following renal ischemic-reperfusion (I/R) injury in rabbits. METHODS: BMSCs were collected and isolated from rabbits. Twenty-eight rabbits were subjected to renal pedicle clamping for 105 min and randomized subsequently into transplantation group and control group. BMSCs or saline were injected into the kidney via the renal artery, respectively. Before and 1, 3, 5, 7, 14, 21, and 28 days after I/R injury the venous blood was collected to measure the serum levels of SCr and BUN, and the renal tissue was sampled for pathological observation. RESULTS: One and 3 days after I/R injury, serum Cr and BUN levels increased significantly to the highest level in both groups. On the 7th day serum Cr and BUN levels in the transplantation group were lower than those in control group and remained so till the end of the experiment. On the 28th day, the levels of serum Cr (90.1+/-11.1 micromol/L) and BUN (8.0+/-1.5 mmol/L) in the transplantation group were significantly lower than those in the control group (135.6+/-32.5 micromol/L and 10.9+/-2.5 mmol/L, respectively, P<0.05). Pathological observation of the renal tissue revealed renal tubular epithelial cell degeneration, necrosis and abscission. CONCLUSION: BMSC transplantation can accelerate renal function repair after acute tubular necrosis resulting from I/R injury, and decrease serum Cr and BUN levels in early stage following the injury.