CTHRC1 promotes colorectal cancer progression by recruiting tumor-associated macrophages via up-regulation of CCL15.

Tumor-associated macrophages (TAMs) represent a key factor in the tumor immune microenvironment (TME), exerting significant influence over tumor migration, invasion, immunosuppressive features, and drug resistance. Collagen triple helix repeat containing 1 (CTHRC1), a 30 KDa protein which was secreted during the tissue-repair process, is highly expressed in several malignant tumors, including colorectal cancer (CRC). Previous studies demonstrated that CTHRC1 expression in TAMs was positively correlated to M2 macrophage polarization and liver metastasis, while our discovery suggesting a novel mechanism that CTHRC1 secreted from cancer cell could indirectly interplay with TAMs. In this study, the high expression level of CTHRC1 was evaluated in CRC based on GEO and TCGA databases. Further, CTHRC1 was detected high in all stages of CRC patients by ELISA and was correlated to poor prognosis. Multispectral imaging of IHC demonstrated that M2 macrophage infiltration was increased accompanied with CTHRC1 enrichment, suggesting that CTHRC1 may have chemotactic effect on macrophages. In vitro, CTHRC1 could have chemotactic ability of macrophage in the presence of HT-29 cell line. Cytokine microarray revealed that CTHRC1 could up-regulate the CCL15 level of HT-29, pathway analysis demonstrated that CTHRC1 could regulate CCL15 by controlling the TGFß activation and Smad phosphorylation level. In vivo, knocking down of CTHRC1 from CT-26 also inhibits tumor formation. In conclusion, CTHRC1 could promote the chemotactic ability of macrophages by up-regulating CCL15 via TGFß/Smad pathway; additionally, a high level of CTHRC1 could promote macrophage's M2 polarization. This discovery may be related to tumor immune tolerance and tumor immunotherapy resistance in CRC. KEY MESSAGES: CTHRC1 promotes CRC progression by up-regulating CCL15 via TGF-ß/Smad pathways to further recruit tumor-associated macrophages. By the means of autocrine or paracrine, CTHRC1 can indeed promote macrophage chemotaxis and enhance the infiltration of macrophages in tumor tissues but in the presence of tumor cells. CAFs were another source of CTHRC1, indicating CTHRC1 can infiltrate tumor islet as well as the stomal and be secreted from both tumor cells and CAFs. This study validated CTHRC1 as a potential immune therapy target CRC.

MRI Radiomic Features: A Potential Biomarker for Progression-Free Survival Prediction of Patients With Locally Advanced Cervical Cancer Undergoing Surgery.

OBJECTIVES: To investigate the prognostic role of radiomic features based on pretreatment MRI in predicting progression-free survival (PFS) of locally advanced cervical cancer (LACC). METHODS: All 181 women with histologically confirmed LACC were randomly divided into the training cohort (n = 126) and the validation cohort (n = 55). For each patient, we extracted radiomic features from whole tumors on sagittal T2WI and axial DWI. The least absolute shrinkage and selection operator (LASSO) algorithm combined with the Cox survival analysis was applied to select features and construct a radiomic score (Rad-score) model. The cutoff value of the Rad-score was used to divide the patients into high- and low-risk groups by the X-tile. Kaplan-Meier analysis and log-rank test were used to assess the prognostic value of the Rad-score. In addition, we totally developed three models, the clinical model, the Rad-score, and the combined nomogram. RESULTS: The Rad-score demonstrated good performance in stratifying patients into high- and low-risk groups of progression in the training (HR = 3.279, 95% CI: 2.865-3.693, p < 0.0001) and validation cohorts (HR = 2.247, 95% CI: 1.735-2.759, p < 0.0001). Otherwise, the combined nomogram, integrating the Rad-score and patient's age, hemoglobin, white blood cell, and lymph vascular space invasion, demonstrated prominent discrimination, yielding an AUC of 0.879 (95% CI, 0.811-0.947) in the training cohort and 0.820 (95% CI, 0.668-0.971) in the validation cohort. The Delong test verified that the combined nomogram showed better performance in estimating PFS than the clinical model and Rad-score in the training cohort (p = 0.038, p = 0.043). CONCLUSION: The radiomics nomogram performed well in individualized PFS estimation for the patients with LACC, which might guide individual treatment decisions.

Role and mechanism underlying FoxO6 in skeletal muscle in vitro and in vivo.

Skeletal muscle atrophy is a common feature of patients suffering with chronic infection and other systemic diseases, including acquired immunodeficiency syndrome, chronic kidney disease and cancer. Therefore, understanding the molecular basis of muscle loss is of importance. The majority of members of the forkhead box O (FoxO) family can induce skeletal muscle atrophy; however, the effect of FoxO6 on skeletal muscle is not completely understood. The present study investigated the role of FoxO6 in vitro and in vivo. Compared with the small interfering RNA (si)­negative control (NC) group, C2C12 cell proliferation (Cell Counting Kit­8 assay), myotube differentiation and myotube production were significantly decreased by FoxO6 knockdown, which was different from the known functions of other FoxO members. The immunofluorescence assay results demonstrated that si­FoxO6 clearly downregulated the expression levels of myosin heavy chain (MyHC) in C2C12 myotubes compared with si­NC. The western blotting results indicated that compared with the si­NC group, FoxO6 knockdown induced C2C12 myotube atrophy by notably downregulating myoblast determination protein 1 (MyoD), mTOR and MyHC expression levels, and by markedly upregulating ubiquitin ligase (atrogin1) and muscle RING­finger protein­1 (MURF1) expression levels. Similarly, in an in vitro model of TNF­α­induced myotube atrophy, the western blotting results indicated that FoxO6 expression levels were decreased, whereas atrogin1, MURF1, FoxO1 and FoxO3a expression levels were increased compared with the control group. Therefore, the results indicated that, unlike FoxO1 or FoxO3a, FoxO6 maintained C2C12 myotubes and protected against atrophy. Consistent with the in vitro data, similar results were observed in vivo. Collectively, the results of the present study suggested that FoxO6 served a critical role in muscle cell metabolism in vitro and in vivo, and might serve as a promising therapeutic target for ameliorating skeletal muscle atrophy.

Ultrasound-based radiomics score: a potential biomarker for the prediction of progression-free survival in ovarian epithelial cancer.

PURPOSE: More than 80% of patients with ovarian epithelial cancer (OEC) show complete remission after initial treatment but eventually experience recurrence of the disease. This study aimed to develop a radiomics signature to identify a new prognostic indicator based on preoperative ultrasound imaging. METHODS: A total of 111 patients with OEC who underwent transvaginal ultrasound before surgery were included. Of these, 76 were divided into the training cohort and 35 into the test cohort. We defined the region of interest (ROI) of the tumor by manually drawing the tumor contour on the ultrasound image of the lesion. The radiomics features were extracted from ultrasound images. The radiomics score (Rad-Score) was constructed using the least absolute shrinkage and selection operator (LASSO) analysis and Cox regression. Combined with the ultrasound radiomics features, significant clinical variables were also used to establish predictive models for 5-year progression-free survival (PFS) prediction. The efficiency of the model was evaluated using the area under the curve (AUC). Kaplan-Meier analysis was used to evaluate the association between the Rad-Score and PFS. RESULTS: The combined model was superior to the clinical and Rad-Score models in estimating 5-year PFS and achieved an AUC of 0.868 (95%CI 0.766-0.971) in the training cohort. The Rad-Score was negatively correlated with prognosis in the training and test cohorts. CONCLUSIONS: The combined model that incorporated both clinical parameters and ultrasound radiomics features achieved a good prognosis in patients with OEC, which might aid clinical decision-making.

Clinical Features, Risk Factors, and Early Prognosis for Wallerian Degeneration in the Descending Pyramidal Tract after Acute Cerebral Infarction.

BACKGROUND: Wallerian degeneration(WD) occurs in the descending pyramidal tract(DPT) after cerebral infarction commonly, but studies of its degree evaluation, influencing factors and effects on nervous function are still limited. OBJECTIVES: The purpose of this study was to describe these findings and estimate their clinical significance. METHODS: In total, 133 patients confirmed acute cerebral infarction and restricted diffusion in the DPT of the cerebral peduncle by MRI scans. These cases were retrospectively reviewed. We describe their clinical characteristics and analyze influence factors of WD, including the timespan from symptom onset to MRI and TOAST classification. Their NIHSS scores at admission and first 7 days NIHSS improvement rate after admission were also analyzed. RESULTS: These patients were divided into three groups by timespan ≤7 days(n = 45),7-14 days(n = 70) and >14 days(n = 18). The mean WD degree (%)of these three groups was 44.41 ± 22.51,52.35 ± 22.61and 44.31 ± 19.35,respectively(p = 0.122).According to the TOAST classification, the mean WD degree(%) of the cardioembolism group(n = 28, 62.80 ± 25.12) was significantly different from both the large-artery atherosclerosis group(n = 73,45.08 ± 20.03,p = 0.000) and the small-vessel occlusion group(n = 23,39.68 ± 16.95,p = 0.000). The mean NIHSS score upon admission of the WD degree≤50% group(n = 82,8.17 ± 5.87) was different from that of the >50% group(n = 51,11.31 ± 7.00)(p = 0.006). However, the mean 7 days NIHSS improvement rate(%) of the WD degree≤50% group(n = 79,11.83 ± 23.76)and >50% group(n = 50,13.40 ± 27.88) was not significantly different(p = 0.733). CONCLUSIONS: Early WD in ischemic stroke patients has a correlation with serious baseline functional defects. Therefore, we should give close attention to imaging change, especially in those with cardioembolism .

Potential role of fecal microbiota in patients with constipation.

BACKGROUND: We evaluated the safety and efficacy of fecal microbiota transplantation (FMT) for chronic functional constipation (CFC) ineffectively treated by conventional constipation medication. METHODS: Thirty-four patients with CFC underwent FMT treatment (three rounds, via gastroscopy). Clinical scales, including the Wexner constipation score as the main index of efficiency, were completed at baseline; after each treatment, and at 2 and 3 months of follow up. Secondary evaluation indices included the self-assessment of constipation symptoms, patient assessment constipation quality-of-life questionnaire, Bristol stool form scale, and Zung's self-rating depression and anxiety scales. Gastrointestinal motility, motilin, gastrin, nitric oxide (NO), and 5-hydroxytryptamine (5-HT) were assessed before and after treatment. Intestinal flora changes were assessed by 16S ribosomal ribonucleic acid (rRNA) sequencing. RESULTS: There were no serious adverse reactions. The clinical cure rate was 73.5% (25/34), clinical remission rate was 14.7% (5/34), and the inefficiency rate was 11.8% (4/34). Clinical scale data indicated that the FMT treatment was effective. Furthermore, FMT treatment promoted intestinal peristalsis, increased gastrointestinal motility, and increased serum NO and 5-HT levels. The 16S rRNA sequencing data indicated that high abundances of Bacteroides, Klebsiella, Megamonas, Erysipelotrichaceae and Epulopiscium may be the cause of constipation, and high abundances of Prevotella, Acidaminococcus and Butyricimonas may be the main factors in curing constipation. CONCLUSION: Treatment with FMT regulates the intestinal microflora and changes the abundance of CFC-associated bacterial flora to improve constipation.

HLA-DPB1 Variant Effect on Hepatitis B Virus Clearance and Liver Cirrhosis Development Among Southwest Chinese Population.

BACKGROUND: Previous studies have shown that genetic variants in HLA-DP genes affect disease progression in hepatitis B virus (HBV) infection. OBJECTIVES: We aimed to evaluate possible association between HLA-DPB1 rs9277534 polymorphism and different clinical complications of hepatitis B virus (HBV) infection. MATERIALS AND METHODS: Snapshot assay was used to investigate the association of rs9277534 polymorphism in 342 patients with persistent HBV infection and 342 age and gender-matched HBV spontaneous clearance controls. Patients were categorized into asymptomatic HBV carriers (AsC, n = 104), chronic hepatitis B (CHB, n = 116), and liver cirrhosis (LC, n = 122) subgroups. RESULTS: There was a significantly higher proportion of the rs9277534 minor allele A in HBV spontaneous clearance control than that in HBV persistent infection group (OR = 0.58, 95%CI = 0.46-0.73, P < 0.0001). Genotypic analysis showed that GA and AA genotypes were associated with HBV spontaneous clearance (GA: OR = 0.56, 95%CI = 0.40-0.79, P = 0.019; AA: OR = 0.24, 95%CI = 0.14-0.44, P < 0.0001). A significant difference was found between AsC and LC groups in the distribution of AA genotype (OR = 9.32, 95%CI = 1.293-67.14, P = 0.027). CONCLUSIONS: Variant at rs9277534 could affect both the spontaneous clearance of HBV infection and progression from asymptomatic HBV carriers to HBV-related liver cirrhosis in Southwest Han Chinese population.

Polymorphisms in CISH gene are associated with persistent hepatitis B virus infection in Han Chinese population.

BACKGROUND AND AIM: Cytokine-inducible SRC homology 2 domain protein (CISH) is the first member of the suppressors of cytokine signaling (SOCS) protein family. An association between multiple CISH polymorphisms and susceptibility to infectious diseases has been reported. This study aimed to investigate the possible association of these single nucleotide polymorphisms (SNPs) in CISH gene with different outcomes of Hepatitis B virus (HBV) infection. METHODS: 1019 unrelated Chinese Han subjects, including 240 persistent asymptomatic HBV carriers, 217 chronic hepatitis B patients, 137 HBV-related liver cirrhosis patients, and 425 cases of spontaneously recovered HBV as controls, were studied. Four SNPs (rs622502, rs2239751, rs414171 and rs6768300) in CISH gene were genotyped with the snapshot technique. Transcriptional activity of the CISH promoter was assayed in vitro using the dual-luciferase reporter assay system. RESULTS: At position rs414171, A allele and AA genotype frequencies were significantly higher in the HBV-resolved group as compared to the persistent HBV infection group. At position rs2239751, TT genotype was further observed in the HBV-resolved group. Using asymptomatic HBV carriers as controls, our results indicated that the rs414171 and rs2239751 polymorphisms were unrelated to HBV progression. The other two SNPs (rs622502 and rs6768300) showed no association with persistent HBV infection. Haplotype analysis revealed that the GGCA haplotype was associated with spontaneous clearance of HBV in this population. Moreover, luciferase activity was significantly higher in the PGL3-Basic-rs414171T construct as compared to the PGL3-Basic-rs414171A construct (p<0.001). CONCLUSION: Two SNPs (rs414171 and rs2239751) in the CISH gene were associated with persistent HBV infection in Han Chinese population, but not with HBV progression.

Cyclin D1 G870A polymorphism and the risk of hepatocellular carcinoma in a Chinese population.

Cyclin D1, encoded by the gene CCND1, is a regulatory protein in the cell cycle transition from G1 phase to S phase. A common polymorphism (G870A) in the exon 4 of CCND1 gene affects splicing of the CCND1 transcript and may cause uncontrollable cellular growth. Therefore, the CCND1 G870A polymorphism may influence an individual's susceptibility to the development of certain tumors. The present study was performed to test the association between G870A polymorphism in the CCND1 gene and hepatocellular carcinoma (HCC) risk in a Chinese population. We extracted the peripheral blood samples from 220 patients with HCC and 220 age- and gender-matched healthy controls. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and direct DNA sequencing were performed to detect the polymorphism. The CCND1 genotype distribution among HCC patients was not significantly different from that among healthy controls (P=0.08). Compared with the wild-type GG genotype, neither the variant AA genotype nor the variant genotypes containing the A allele were associated with risk of HCC. However, stratification analysis by HBV carrier status revealed that the variant genotypes containing the A allele were associated with a significantly increased risk of HCC among HBsAg-positive individuals (adjusted OR=3.87; 95 % CI=1.12, 13.30). These results suggest that the CCND1 G870A polymorphism may increase the risk of HBV-related HCC in the Chinese population.

[Significance of the double mutations of C1673T/C1799G in HBV C promoter].

OBJECTIVE: To explore the biological and clinical significance of the double mutations of C1673T/C1799G in HBV C promoter (CP). METHODS: Totally 136 patients were enrolled, including 25 asymptomatic carriers (AsC), 38 patients with chronic hepatitis B (CHB), 24 patients with chronic severe hepatitis B (CSHB), 36 cases with liver cirrhosis (LC) and 13 cases with hepatocellular carcinoma (HCC). HBV subgenotypes and mutations in CP of all samples were determined by nested-PCR and direct nucleotide sequence analysis. The C to T mutation at nucleotide 1673 and C to G at nucleotide 1799 were analyzed in different subgenotypes, and the relationships of C1673T/C1799G double mutations with HBV replication, the expression of HBeAg, and with the severity of liver disease after chronic HBV infection were studied. RESULTS: Of the 136 patients, 110 were subgenotype Ba, 1 was Bj, 7 were C1, and 18 were C2. C1673T/C1799G double mutations in Ba were determined in 106 (96. 4%) samples, which was significantly higher than in C1 (14.3%) and C2 (12.5%) subgenotype (P < 0.0001). In contrast to non-mutation group, HBV DNA content in mutation group had no significant difference (P > 0.05). The prevalence of the mutation was lower in HBeAg positive patients (71.4%) than in HBeAg negative patients (87.5%) (P < 0.05). The frequencies of the double mutations were not significantly different among ASC, CHB, CSHB, LC and HCC groups (P > 0.05). CONCLUSION: In Ba subgenotype, double mutations of C1673T/C1799G is much popular than in C1 and C2; the mutation has no effect on HBV replication, and may not be associated with the outcome of chronic HBV infection.

[Effects of human urinary tissue kallikreins on vasodilation of basilar artery in rabbits with symptomatic cerebral vasospasm].

OBJECTIVE: To evaluate the effects of urinary kallidinogenase on subarachnoid hemorrhage (SAH) in rabbits. METHODS: Rabbits symptomatic cerebral vasospasm model was built though Endo method, among the 40 rabbits, 8 died or had severe nervous system syndrome, the other 32 were randomly divided into 4 groups:group A, control group, injection of normal saline to the cisterna magna;group B, subarachnoid hemorrhage;group C, injection of human urinary tissue kallikreins;group D, treated with Nimodipine. The behavior scores, neurological scores and cerebral angiography changes were observed. RESULTS: Food intake obviously decreased and neurological deficit were seen in group B, while which were attenuated in group C and group D, and group A was normal. Comparing the diameter of basilar artery was (1.9 +/- 0.3) mm before SAH, the diameter of group B 4 d later was (1.5 +/- 0.3) mm, 7 d later (1.4 +/- 0.3) mm, the difference was significant (P < 0.05). Comparing with group C on the day 4th and 7th, the diameters of basilar artery were significantly different (P < 0.001). Comparing with group D on the day 4th, 7th and 14th, there was no obvious improvement. CONCLUSION: Urinary kallidinogenase and Nimodipine can obviously alleviate symptomatic cerebral vasospasm in rabbits remarkably, but the former's effect of attenuating vasospasm is better than that of Nimodipine.

[Effects of human tissue kallikrein on cerebral vasospasm: experiment with rabbits].

OBJECTIVE: To study the effects of human tissue kallikrein (HTK) on symptomatic cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH). METHODS: Forty rabbits underwent occlusion of bilateral carotid. Two weeks later the 28 surviving rabbits were randomly divided into to 4 groups: shamed-operation group (n = 8) undergoing injection of normal saline into the cisterna magna on day 1 and day 3, SAH group (n = 6) undergoing injection of nonheparinized autologous arterial blood into the cisterna magna, HTK therapy group (n = 6) undergoing blood injection into the cisterna magna and then injection of HTK via ear marginal vein daily for 3 days, and nimodipine (ND) therapy group (n = 6) undergoing blood injection into the cisterna magna and then injection of ND via ear marginal vein. 3-dimension-CT angiography (3-D CTA) was used to measure the basilar artery diameter on D(0) and D(5). On D(6) the rabbits were killed with their basilar arteries taken out to undergo light microscopic examination. RESULTS: Blood could be seen in the basis cephalic of the 3 groups undergoing blood injection. 3-D CTA showed that arteriospasm was seen in the SAH and ND groups but not in the HTK group. Microscopy showed obvious pathological changes in basilar artery in the SAH and ND groups but not in the HTK group. CONCLUSION: HTK given early after SAH effectively alleviates the symptomatic cerebral vasospasm.