Prognoses of Patients Treated With Surgical Therapy Versus Continuation of Local-Plus-Systemic Therapy Following Successful Down-Staging of Intermediate-Advanced Hepatocellular Carcinoma: A Multicenter Real-World Study.

BACKGROUND: The difference in the prognoses between treatment with surgical therapy and continuation of local-plus-systemic therapy following successful down-staging of intermediate-advanced hepatocellular carcinoma (HCC) remains unclear. METHODS: Data of 405 patients with intermediate-advanced HCC treated at 30 hospitals across China from January 2017 to July 2022 were retrospectively reviewed. All patients received local-plus-systemic therapy and were divided into the surgical (n = 100) and nonsurgical groups (n = 305) according to whether they received surgical therapy. The differences between long-term prognoses of the 2 groups were compared. Subgroup analysis was performed in 173 HCC patients who met the criteria for surgical resection following down-staging. RESULTS: Multivariable analysis of all patients showed that surgical therapy, hazard ratio (HR): 0.289, 95% confidence interval, CI, 0.136-0.613) was a protective factor for overall survival (OS), but not for event-free survival (EFS). Multivariable analysis of 173 intermediate-advanced HCC patients who met the criteria for surgical resection after conversion therapy showed that surgical therapy (HR: 0.282, 95% CI, 0.121-0.655) was a protective factor for OS, but not for EFS. Similar results were obtained after propensity score matching. For patients with Barcelona Clinic Liver Cancer stage B (HR: 0.171, 95% CI, 0.039-0.751) and C (HR: 0.269, 95% CI, 0.085-0.854), surgical therapy was also a protective factor for OS. CONCLUSIONS: Overall, for patients with intermediate-advanced HCC who underwent local-plus-systemic therapies, surgical therapy is a protective factor for long-term prognosis and can prolong OS, and for those who met the surgical resection criteria after conversion therapy, surgical therapy is recommended.

Clinical study on conversion therapy of hepatocellular carcinoma - summary and comparison of clinical data from a single center of consecutive four years.

AIM: The purpose of this study was to interpret real-world clinical data to analyze the surgical safety and survival outcomes of patients with initial unresectable hepatocellular carcinoma (uHCC) after conversion therapy. METHODS: A retrospective analysis was performed on 2984 hepatocellular carcinoma (HCC) patients hospitalized in Shandong Cancer Hospital Affiliated to Shandong First Medical University from June 1st, 2019 to June 1st, 2023. Clinicopathological features, response to systemic and/or loco-regional treatments, surgical resection rate after conversion therapy, surgical safety, and postoperative recurrence were analyzed. RESULTS: A total of 38 patients were successfully converted to obtain surgical resection. 35 patients underwent radical resection. A high objective response rate (ORR) (52.6% under RECIST v1.1 and 78.9% under mRECIST criteria) was observed in patients under conversion therapy, and the disease control rate (DCR) was 100%. Pathologic complete response (pCR) was 42.9%. Treatment-related adverse events (TRAEs) of any grade were observed in 37 patients (97.4%). Safety of conversion or direct surgery continues to improve. The median follow-up time was 19.3 months. The 1-year Disease-free survival (DFS) rate of patients with direct surgery and patients with conversion surgery were 91.4% and 86.8%, respectively. CONCLUSIONS: With conversion therapy, a small percentage (1.81%) of uHCC patients are likely to be converted to radical resection. Local combined systemic therapy is a relatively safe and effective conversion therapy, and the safety of surgery is gradually improved after successful conversion. Preliminary follow-up data showed satisfactory survival benefits for patients undergoing conversion surgery. TRIAL REGISTRATION: This was a retrospective study and it did not interfere with treatment decisions.

Real-world practice of conversion surgery for unresectable hepatocellular carcinoma - a single center data of 26 consecutive patients.

AIM: To understand the proportion of uHCC (unresectable hepatocellular carcinoma) patients who achieve successful conversion resection in a high-volume setting with state of the art treatment options. METHODS: We retrospectively reviewed all HCC patients hospitalized to our center from June 1st, 2019 to June 1st, 2022. Conversion rate, clinicopathological features, response to systemic and/or loco-regional therapy and surgical outcomes were analyzed. RESULTS: A total of 1,904 HCC patients were identified, with 1672 patients receiving anti-HCC treatment. 328 patients were considered up-front resectable. Of the remaining 1344 uHCC patients, 311 received loco-regional treatment, 224 received systemic treatment, and the remainder (809) received combination systemic plus loco-regional treatment. Following treatment, one patient from the systemic group and 25 patients from the combination group were considered to have resectable disease. A high objective response rate (ORR) was observed in these converted patients (42.3% under RECIST v1.1 and 76.9% under mRECIST criteria). The disease control rate (DCR) reached 100%. 23 patients underwent curative hepatectomy. Major post-operative morbidity was equivalent in the both groups (P=0.76). Pathologic complete response (pCR) was 39.1%. During conversion treatment, grade 3 or higher treatment-related adverse events (TRAEs) were observed in 50% of patients. The median follow-up time was 12.9 months (range, 3.9~40.6) from index diagnosis and 11.4 months (range, 0.9~26.9) from resection. Three patients experienced disease recurrence following conversion surgery. CONCLUSIONS: By intensive treatment, a small sub-group of uHCC patients (2%) may potentially be converted to curative resection. Loco-regional combined with systemic modality was relative safe and effective in the conversion therapy. Short-term outcomes are encouraging, but long-term follow-up in a larger patient population are required to fully understand the utility of this approach.

Physicochemical properties and energy content of yellow dent corn from different climatic origins in growing pigs.

OBJECTIVE: The objective of this study was to determine the digestible energy (DE) and metabolizable energy (ME) of yellow dent corn sourced from different meteorological origins fed to growing pigs and develop equations to predict the DE and ME of yellow dent corn from southwestern China. METHODS: Sixty crossbred barrows were allotted to 20 treatments in a triplicate 20×2 incomplete Latin square design with 3 replicated pigs per dietary treatment during 2 consecutive periods. Each period lasted for 12 days, and total feces and urine during the last 5 days of each period were collected to calculate the energy contents. RESULTS: On dry matter (DM) basis, the DE and ME in 20 corn grain samples ranged from 15.38 to 16.78 MJ/kg and from 14.93 to 16.16 MJ/kg, respectively. Selected best-fit prediction equations for DE and ME (MJ/kg DM basis) for yellow dent corn (n = 16) sourced from southwestern China were as follows: DE = 28.58-(0.12×% hemicellulose)+(0.35×% ether extract)-(0.83×MJ/kg gross energy)+(0.20×% crude protein)+(0.49×% ash); ME = 30.42- (0.11×% hemicellulose)+(0.31×% ether extract)-(0.81×MJ/kg gross energy). CONCLUSION: Our results indicated that the chemical compositions, but not the meteorological conditions or physical characteristics could explain the variation of energy contents in yellow dent corn sourced from southwestern China fed to growing pigs.

Metabolizable energy requirement for maintenance estimated by regression analysis of body weight gain or metabolizable energy intake in growing pigs

Objective: Feed energy required for pigs is first prioritized to meet maintenance costs. Additional energy intake in excess of the energy requirement for maintenance is retained as protein and fat in the body, leading to weight gain. The objective of this study was to estimate the ME requirements for maintenance (MEm) by regressing body weight gain against ME intake (MEI) in growing pigs. Methods: Thirty-six growing pigs (26.3 ± 1.7 kg) were allotted to 1 of 6 treatments with 6 replicates per treatment in a randomized complete block design. Treatments were 6 feeding levels which were calculated as 50, 60, 70, 80, 90 or 100% of the estimated ad libitum MEI (2,400 kJ/kg BW0.60·d). All pigs were individually housed in metabolism crates for 30 d and weighed every 5 d. Moreover, each pig from each treatment was placed in the open-circuit respiration chambers to measure heat production (HP) and energy retained as protein (REp) and fat (REf) every 5 d. Serum biochemical parameters of pigs were analyzed at the end of the experiment. Results: The ADG and HP as well as the REp and REf linearly increased with increasing feed intake (p < 0.010). ß-hydroxybutyrate (BHBA) concentration of serum tended to increase with increasing feed intake (p = 0.080). The regression equations of MEI on ADG were MEI, kJ/kg BW0.60·d = 1.88 × ADG, g/d + 782 (R2=0.86) and MEm was estimated at 782 kJ/kg BW0.60·d. Protein retention of growing pigs would be positive while REf would be negative at this feeding level via regression equations of REp and REf on MEI. Conclusion: The MEm was estimated at 782 kJ/kg BW0.60·d in current experiment. Furthermore, growing pigs will deposit protein and oxidize fat if provided feed at the estimated maintenance level.

Net energy content of rice bran, corn germ meal, corn gluten feed, peanut meal, and sunflower meal in growing pigs.

OBJECTIVE: The objective of this experiment was to determine the net energy (NE) content of full-fat rice bran (FFRB), corn germ meal (CGM), corn gluten feed (CGF), solvent-extracted peanut meal (PNM), and dehulled sunflower meal (SFM) fed to growing pigs using indirect calorimetry or published prediction equations. METHODS: Twelve growing barrows with an average initial body weight (BW) of 32.4±3.3 kg were allotted to a replicated 3×6 Youden square design with 3 successive periods and 6 diets. During each period, pigs were individually housed in metabolism crates for 16 d, which included 7 days for adaptation. On d 8, the pigs were transferred to the respiration chambers and fed one of the 6 diets at 2.0 MJ metabolizable energy (ME)/kg BW0.6/d. Total feces and urine were collected and daily heat production was measured from d 9 to d 13. On d 14 and d15, pigs were fed at their maintenance energy requirement level. On the last day pigs were fasted and fasting heat production was measured. RESULTS: The NE of FFRB, CGM, CGF, PNM, and SFM measured by indirect calorimetry method was 12.33, 8.75, 7.51, 10.79, and 6.49 MJ/kg dry matter (DM), respectively. The NE/ME ratios ranged from 67.2% (SFM) to 78.5% (CGF). The NE values for the 5 ingredients calculated according to the prediction equations were 12.22, 8.55, 6.79, 10.51, and 6.17 MJ/kg DM, respectively. CONCLUSION: The NE values were the highest for FFRB and PNM and the lowest in the corn co-products and SFM. The average NE of the 5 ingredients measured by indirect calorimetry method in the current study was greater than values predicted from NE prediction equations (0.32 MJ/kg DM).

Evaluation of available energy and total tract digestibility of acid-hydrolyzed ether extract of cottonseed oil for growing pigs by the difference and regression methods.

OBJECTIVE: The objective of this study was to determine the effect of inclusion level on the digestible energy (DE), metabolizable energy (ME), and total tract digestibility of acid-hydrolyzed ether extract (AEE) of cottonseed oil when fed to growing pigs. METHODS: Forty-two barrows (initial body weight = 35.51±2.01 kg) were randomly allotted to a completely randomized design with a corn-soybean meal basal diet, five levels of cottonseed oil (2%, 4%, 6%, 8%, and 10%) and a 10% soybean oil diet. Each diet was replicated six times with one pig per replicate. The experiment lasted 19 days, 7 d for cage adaptation, 7 d for diets adaptation and last 5 d for feces and urine collection. The energy values and apparent total tract digestibility (ATTD) of cottonseed oil and soybean oil were calculated by the difference method, and regression equations were established to predict the energy values of cottonseed oil. The apparent digested fat of the entire intestinal tract was also regressed against dietary fat intake to determine the true total tract digestibility (TTTD) and endogenous loss of fat for cottonseed oil. RESULTS: The results showed that the DE and ME contents of cottonseed oil were not different as the inclusion level increased. The DE and ME values determined by the regression equation were 36.28 MJ/kg and 34.96 MJ/kg, respectively, and the values were similar to the mean DE and ME values calculated by the difference method (36.18 and 35.56 MJ/kg, respectively). The ATTD of cottonseed oil was also not affected by the inclusion level of cottonseed oil, and the TTTD and EFL determined by the regression method were 92.40% and 13.83 g/kg of dry matter intake for corn-soybean basal diet. The DE, ME, and ATTD of AEE in soybean oil determined by the difference method were 35.70 MJ/kg, 35.20 MJ/kg and 92.31%, respectively. There were no differences in the DE, ME, and ATTD between cottonseed oil and soybean oil, although the ratio of unsaturated to saturated fatty acids for soybean oil was higher than for cottonseed oil. CONCLUSION: The DE, ME, and ATTD values of cottonseed oil were not affected by its dietary inclusion level. The energy values of cottonseed oil determined by the difference and regression methods were similar. Furthermore, the ratio of unsaturated to saturated fatty acid for oils was not the decisive factor to influence the energy values and ATTD of oils.

Oligosaccharides are a key factor in prediction of amino acid digestibility in soybean meal of different origins when fed to growing pigs.

OBJECTIVE: The objective of this experiment was to determine apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of crude protein (CP) and amino acid (AA) in 15 sources of soybean meal (SBM) produced from soybeans from different countries and subsequently to establish equations for predicting the AID and SID in SBM based on their chemical composition. METHODS: Eighteen barrows (57.9±6.1 kg) fitted with a simple T-cannula were allotted into three 6×6 Latin square designs. Each period comprised a 6-d adaption period followed by a 2-d collection of ileal digesta. The 15 test diets included SBM as a sole source of AA in the diet. Another nitrogen-free diet was used to measure basal endogenous losses of CP and AA. Chromic oxide (0.3%) was used as an inert marker in each diet. RESULTS: The AID of lysine in SBM from China and USA tended to be greater than in SBM from Brazil (p<0.10). The SID of valine and proline in SBM from China was greater than in SBM from Brazil (p<0.05). The SID of lysine, threonine, cysteine and glycine in SBM from China tended to be greater than in SBM from Brazil (p<0.10). From a stepwise regression analysis, a series of AID and SID prediction equations were generated. The best fit equations for lysine in SBM were: AID lysine = 1.16 sucrose-1.81 raffinose+82.10 (R2 = 0.69, p<0.01) and SID lysine = 1.14 sucrose-1.93 raffinose-0.99 ether extract (EE)+85.26 (R2 = 0.77, p<0.01). CONCLUSION: It was concluded that under the conditions of this experiment, the oligosaccharides (such as sucrose and raffinose) can be used to predict the AID and SID of AA in SBM with reasonable accuracy.

Prediction of digestible and metabolisable energy in soybean meals produced from soybeans of different origins fed to growing pigs.

The objective of this experiment was to determine the digestible energy (DE) and metabolisable energy (ME) in 22 sources of soybean meal (SBM) produced from soybeans from different countries and subsequently to establish equations for predicting the DE and ME in SBM based on their chemical composition. The 22 sources of SBM were all processed in Chinese crushing plants, but the soybeans used originated from China (n=6), the US (n=6), Brazil (n=7) or Argentina (n=3). The basal diet was a corn-based diet and 22 additional diets were formulated by mixing corn and 24.3% of each source of SBM. The average DE and ME in SBM from China, the US, Brazil and Argentina were 15.73, 15.93, 15.64 and 15.90 MJ/kg and 15.10, 15.31, 14.97 and 15.42 MJ/kg, respectively, and no differences among countries were observed. From a stepwise regression analysis, a series of DE and ME prediction equations were generated. The best-fit equations for SBM were DE=38.44-0.43 crude fibre -0.98 gross energy +0.11 acid detergent fibre (R2=0.67, p<0.01) and ME=2.74+0.97 DE -0.06 crude protein (R2=0.79, p<0.01). In conclusion, there were no differences in the DE and ME of SBM among the different soybean sources used in this experiment. The DE and ME of SBM of different origin can be predicted based on their chemical composition when fed to growing pigs.

The Effect of Inclusion Level of Soybean Oil and Palm Oil on Their Digestible and Metabolizable Energy Content Determined with the Difference and Regression Method When Fed to Growing Pigs.

This experiment was conducted to determine the effects of inclusion level of soybean oil (SO) and palm oil (PO) on their digestible and metabolism energy (DE and ME) contents when fed to growing pigs by difference and regression method. Sixty-six crossbred growing barrows (Duroc×Landrace×Yorkshire and weighing 38.1±2.4 kg) were randomly allotted to a 2×5 factorial arrangement involving 2 lipid sources (SO and PO), and 5 levels of lipid (2%, 4%, 6%, 8%, and 10%) as well as a basal diet composed of corn and soybean meal. The barrows were housed in individual metabolism crates to facilitate separate collection of feces and urine, and were fed the assigned test diets at 4% of initial body weight per day. A 5-d total collection of feces and urine followed a 7-d diet adaptation period. The results showed that the DE and ME contents of SO and PO determined by the difference method were not affected by inclusion level. The DE and ME determined by the regression method for SO were greater compared with the corresponding respective values for PO (DE: 37.07, ME: 36.79 MJ/kg for SO; DE: 34.11, ME: 33.84 MJ/kg for PO, respectively). These values were close to the DE and ME values determined by the difference method at the 10% inclusion level (DE: 37.31, ME: 36.83 MJ/kg for SO; DE: 34.62, ME: 33.47 MJ/kg for PO, respectively). A similar response for the apparent total tract digestibility of acid-hydrolyzed ether extract (AEE) in lipids was observed. The true total tract digestibility of AEE in SO was significantly (p<0.05) greater than that for PO (97.5% and 91.1%, respectively). In conclusion, the DE and ME contents of lipid was not affected by its inclusion level. The difference method can substitute the regression method to determine the DE and ME contents in lipids when the inclusion level is 10%.

[Current treatment and progress of hepatocellular carcinoma with portal vein tumor thrombus].

Portal venous tumor emboli is one of the important factors that affect the prognosis of patients with hepatocellular carcinoma (HCC). There are different characters of surgery, interventional therapy, radiation therapy, treatment of local damage, targeted therapy in the treatment of portal venous tumor emboli, but the effects are not ideal. Scholars at home and abroad are exploring a variety of treatment patterns in the treatment of portal venous tumor emboli, to achieve better effect.Predominantly surgical comprehensive treatment can excision of tumor and tumor emboli, improve liver function, improve the quality of survival, prolong survival time. But the operation risk is big. Mainly non-surgical treatment has advantages of little trauma, less risk, and better local control of tumor emboli. But the efficiency, effect, alleviate period is ineffective. Therefore, the best treatment mode should be explored according to the different tumor emboli parting.

Effect of experimental methodology on fasting heat production and the net energy content of corn and soybean meal fed to growing pigs.

The aim of the three experiments was to evaluated methods to predict fasting heat production (FHP) and to compare methods to determine the net energy (NE) of corn and soybean meal (SBM) fed to growing pigs. To estimate heat production (HP), pigs were housed in respiratory chambers for all experiments. In Experiment 1, six barrows (43.0 ± 1.4 kg body weight [BW]) were fed a Corn-SBM diet for 20 d. The experimental design consisted of following periods: 7 d adaptation, 5 d ad libitum feeding, 3 d feeding at 2 × metabolisable energy (ME) for maintenance (MEm), 3 d feeding at 1 × MEm and 2 d fasting. The FHP was calculated by extrapolating HP measured at the different feeding levels to zero ME intake. The daily FHP [per kg BW(0)(.6)] determined directly after fasting for 24 h and using the regression method was 774 kJ and 694 kJ, respectively. In Experiment 2, 18 barrows (34.3 ± 1.1 kg BW) were randomly allotted to three diets: Diet 1 contained 97.5% corn (direct NE determination of corn); diets 2 and 3 contained 25 % and 15% SBM at the expense of corn, respectively, and were used to calculate the NE of corn by difference. The NE of corn determined directly (13.21 MJ/kg DM) and by difference (13.69 MJ/kg DM) was not different. In Experiment 3, 24 barrows (36.2 ± 1.4 kg BW) were randomly allotted to four diets to determine the effects of different basal diets on the NE content of SBM. The diets were: Basal diet 1 (97.5% corn), Test diet 1 (15% SBM at the expense of corn), Basal diet 2 (contained 72.5% corn and 25% SBM) and Test diet 2 (58% corn and 39.5% SBM). These diets were used to determine the NE of SBM using the Corn-basal diet or the Corn-SBM-basal diet, respectively. It was shown that the estimated NE of SBM did not depend on the used diet (10.04 MJ/kg and 10.62 MJ/kg DM for Basal diet 1 and 2, respectively). In summary, using the regression method to determine FHP results in lower FHP than the fasting method. There was no difference observed in the NE of corn determined directly or by difference, and different basal diets did not affect the NE of SBM.

Achieving complete remission in metastatic hepatocellular carcinoma with sintilimab plus sorafenib therapy followed by hepatic resection: a case report.

Background: The synergistic effectiveness of combining immune checkpoint inhibitors with targeted therapies has shown promise in improving the conversion rate for unresectable hepatocellular carcinoma (HCC) patients to a potentially resectable status. However, the efficacy of this approach in the context of HCC with extrahepatic metastasis remains to be conclusively determined. Case presentation: We report a rare case of advanced HCC with extrahepatic metastasis who achieved long-term survival by a combination of systemic therapy (sintilimab and sorafenib) followed by laparoscopic hepatectomy. A 63-year-old man presented at our hospital with discomfort on the right side of his waist. An enlarged right hepatic lobe mass was subsequently revealed by CT scan. The patient's medical history, including a prior infection with hepatitis B virus, cirrhosis of the liver and an alpha-fetoprotein (AFP) level measuring 41.28 ng/ml substantiated the clinical diagnosis of HCC. On October 30th, 2019, the patient received 200 mg sintilimab intravenously (q3w) plus 200-400 mg BID sorafenib orally, along with antiviral therapy. After six cycles, his disease achieved partial response (PR). On April 26th, 2021, He underwent a laparoscopic hepatectomy. The patient achieved a sustained period of no evidence of disease for 2.5 years and with drug-free survival for 2 years after the resection. His current overall survival is estimated at approximately 4 years. Conclusions: This case highlights the potential of combining sintilimab and sorafenib in transforming HCC with extrahepatic metastasis into a condition amenable to surgical resection, suggesting that this treatment approach, followed by surgery, may lead to complete remission.

Analysis of efficacy and safety for the combination of tislelizumab and regorafenib in advanced hepatocellular carcinoma: A prospective clinical study.

BACKGROUNDS: Programmed death receptor 1 (PD-1) monoclonal antibody has been approved for the first and second-line treatments of hepatocellular carcinoma (HCC). This study aimed to evaluate the efficacy and safety of tislelizumab + regorafenib as a second-line treatment option for advanced HCC. METHODS: Treatment-related adverse events (TRAEs) were the primary endpoints in this clinical trial comprising 28 patients with advanced HCC. The secondary endpoints included objective response rate (ORR), disease control rate (DCR), and progression-free survival (PFS). RESULTS: According to the mRECIST 1.1 evaluation criteria, the ORR was 28.6%. Complete and partial response were observed in 3 and 5 patients, respectively; stable disease was observed in 12 patients (DCR, 71.4%). The median PFS was 6.4 months. The incidence of grade 1-2 and 3-4 TRAEs was 57.1% and 39.3%, respectively. CONCLUSION: This study suggests that tislelizumab + regorafenib can be used as a second-line treatment for advanced HCC.

Neoadjuvant tislelizumab plus stereotactic body radiotherapy and adjuvant tislelizumab in early-stage resectable hepatocellular carcinoma: the Notable-HCC phase 1b trial.

Notable-HCC (NCT05185531) is a phase 1b trial, aiming to evaluate the safety and preliminary effectiveness of neoadjuvant PD-1 blockade plus stereotactic body radiotherapy (SBRT) in early-stage resectable hepatocellular carcinoma (HCC). Twenty patients with HCC of BCLC stage 0-A received 3 × Gy SBRT and two cycles of tislelizumab, an anti-PD-1 monoclonal antibody before the curative HCC resection. Primary endpoints were the surgery delay, radiographic and pathological tumor response after the neoadjuvant therapy, safety and tolerability. During the neoadjuvant therapy, treatment-related adverse events (TRAEs) of grade 1-2 occurred in all 20 patients (100%), eight patients (40%) had grade 3 TRAEs, no grade 4 to 5 TRAE occurred, and all resolved without corticosteroids treatment. Per mRECIST, the objective response rate was 63.2% (12/19), with 3 complete response; the disease control rate was 100%. Two (10.5%) patients achieved complete pathological response. No surgery delay occurred. The neoadjuvant therapy did not increase the surgical difficulty or the incidence of complications. Secondary endpoints of disease-free survival and overall survival were not mature at the time of the analysis. Our pilot trial shows that neoadjuvant therapy with anti-PD-1 + SBRT is safe and promotes tumor responses in early-stage resectable HCC.

Characterization of disease-associated microbiota in hepatocellular carcinoma.

Aims: This study aimed to investigate the differences in the composition of microbial communities and related functions in hepatocellular carcinoma (HCC) tumours and matched normal tissues were investigated. Methods and Material: Tumour tissues and matched normal samples were collected from 30 HCC patients. Genomic DNA was collected and subjected to sequencing of the V3 + V4 region of the 16S rRNA gene. The microbial community profiles and metabolic pathway predictions of the different groups were characterized and compared. Results: Tumour and adjacent tissues had similar microbiota compositions but differed in abundance. Proteobacteria and Firmicutes abundance decreased and Cyanobacteria and Acidobacteria abundance increased in the tumour tissue. The microbial community diversity was higher in the tumour tissues than in adjacent samples, with potentially more dominant taxa in the adjacent tissues, including Firmicutes, Proteobacteria, and Actinobacteria. Acidobacteria, Cyanobacteria, and Chloroflexi were the dominant microbes in tumour tissues. A total of 46 metabolic pathways were identified. Global and overview maps were the most abundant pathways, followed by carbohydrate metabolism, energy metabolism, metabolism of cofactors and vitamins, and membrane transport. The top 50 most highly correlated microbial genera included Klebsiella, Rhodococcus, Ochrobactrum, and Azoarcus. Fonticella, Haloimpatiens, Brevibacterium, and Acidothermus were positively correlated with other microbial genera. The microbiota of adjacent tissues was more robust in the network analysis. Conclusions: This study revealed differences in microbial composition between HCC tumour tissues and normal tissues and differences in microbial abundance associated with different metabolic functions. Cyanobacteria, Proteobacteria, and Actinobacteria may play important roles in HCC.

Tumour-derived exosomal lncRNA SNHG16 induces telocytes to promote metastasis of hepatocellular carcinoma via the miR-942-3p/MMP9 axis.

BACKGROUND: Hepatocellular carcinoma (HCC) cell-derived exosomal LncRNA SNHG16 is highly expressed and associated with poor overall survival of patients. Telocytes (TCs), as novel interstitial cells, have been reported to promote HCC metastasis. Therefore, in our study, we investigated whether a molecular interaction occurred between exosomal LncSNHG16 and TCs in the tumor microenvironment. METHODS: LncSNHG16 expression in HCC tissues and cell lines was measured, and bioinformatics analysis was performed. Exosomes were isolated and purified from HCC cells with LncSNHG16 overexpression/knockdown vectors and cocultured with TCs. Then, markers of the LncSNHG16/miR-942-3p/MMP9 axis were tested in TCs. Transwell assays and cell wound healing assays were designed to examine the invasion and migration of HCC cells after coincubation with TCs. RNA immunoprecipitation (RIP) assays and dual-luciferase gene reporter assays were performed to verify the binding effect of LncSNHG16, miR-942-3p, and MMP9 mRNA. In vivo, experimental animal models were established to confirm the effect of exosomal LncSNHG16-induced MMP9 expression on HCC metastasis. RESULTS: Exosomal LncSNHG16 was phagocytized by TCs and downregulated miR-942-3p, which induced targeted MMP9 upregulation, and it had specific binding sites with miR-942-3p in TCs to facilitate the migration of HCC cells in vitro and in vivo. Exosomal LncSNHG16 was found to act as a competing endogenous RNA of the miR-942-3p/MMP9 axis in TCs. CONCLUSION: Tumour-derived exosomal LncSNHG16 modulates MMP9 via competitively binding to miR-942-3p in TCs, thus promoting the metastasis of HCC.

Exosomal miR-200b-3p induce macrophage polarization by regulating transcriptional repressor ZEB1 in hepatocellular carcinoma.

PURPOSE: Accumulating evidence has elucidated that the interaction between cancer cells and M2 macrophages plays an important role in the tumorigenesis of hepatocellular carcinoma (HCC). However, the mechanism connecting tumor-derived exosomes, M2 polarization of macrophages, and liver metastasis remain unclear. Therefore, it is necessary to explore their influence on the tumor microenvironment of HCC. METHODS: Transmission electron microscopy, nanometer particle testing, and special biomarker analysis were utilized to characterize exosomes, while the differential expression of microRNAs was evaluated using high-throughput sequencing technology. The functions of miR-200b-3p exosomes were confirmed using in vitro and in vivo assays. The interactions between microRNAs and ZEB1 as well as cancer cells and macrophages were measured using RNA pull-down and luciferase gene reporter assays. RESULTS: Using in silico analysis, we identified high levels of miR-200b-3p exosome expression in patients with HCC, particularly with relapsed HCC. We demonstrated that HCC cell-derived miR-200b-3p exosomes were internalized by M0 macrophages and induced M2 polarization by downregulating ZEB1 and upregulating interleukin-4. As a result, the JAK/STAT signaling pathway was activated in M2 macrophages, leading to increased PIM1 and VEGFα expression. These cell factors accelerated the proliferation and metastasis of HCC, resulting in a feedback loop between HCC cells and M2 macrophages. CONCLUSION: The study illustrates that HCC cell-derived miR-200b-3p exosomes facilitate the proliferation and polarization of macrophages by modulating cytokine secretion and the JAK/STAT signaling pathway, leading to the metastasis of HCC. These findings demonstrate the existence of a novel feedback loop between cancer cells and immune cells in the tumor microenvironment, presenting a new concept in cancer research.

The hypoxia-inducible factor 1 inhibitor LW6 mediates the HIF-1α/PD-L1 axis and suppresses tumor growth of hepatocellular carcinoma in vitro and in vivo.

The low survival rate of hepatocellular carcinoma (HCC) remains a major challenge for clinicians and patients, and its progression may be related to hypoxia-inducible factor (HIF) and PD-L1. LW6 is a drug that inhibits hypoxia by reducing HIF-1α accumulation and gene transcriptional activity. However, its effect and regulatory mechanism in HCC remain to be revealed, especially under hypoxic conditions. The HIF-1α and PD-L1 expression in HCC specimens and paracarcinoma tissues was evaluated by a tissue microarray (TMA). The effects of LW6 were evaluated by cell viability, colony formation, and Transwell assays and xenografted nude mice. Cell cycle and apoptosis of HCC cells were detected by flow cytometry. The effects of LW6 on HIF-1α signaling and its targets PD-L1 and VEGF were evaluated through qRT-PCR, Western blots, Cell transfection, Transwell migration and invasion assays, immunohistochemistry, immunofluorescence and luciferase assays. In this study, we found that LW6 had antiproliferative effects on HCC and promoted HCC cell apoptosis, inhibited their migration and invasion, and affected their cell cycle. LW6 dramatically decreased HIF-1α expression through the VHL-dependent proteasome system pathway, inhibited HIF-1α transcriptional activation, and reduced PD-L1 expression by inhibiting EGFR pathway activation. These results suggest that LW6 can promote apoptosis of HCC cells by inhibiting HIF-1α, inhibit tumor angiogenesis, and downregulate the expression of PD-L1, which is an effective choice for the treatment of HCC. Moreover, inhibiting the hypoxic microenvironment combined with immunotherapy is expected to be a potentially effective strategy.

Protocol of notable-HCC: a phase Ib study of neoadjuvant tislelizumab with stereotactic body radiotherapy in patients with resectable hepatocellular carcinoma.

INTRODUCTION: Liver resection is the mainstay of curative-intent treatment for hepatocellular carcinoma (HCC), but the postoperative 5-year recurrence rate reaches 70%, and there are no adjuvant or neoadjuvant therapies recommended by major HCC guidelines that can reduce the risk of recurrence. In the recent decade, significant progress has been achieved in the systemic treatment of HCC, mainly from immune checkpoint inhibitors (ICIs) and targeted therapy. In other malignancies, ICIs in the neoadjuvant setting have shown better outcomes than in the adjuvant setting. On the other hand, the addition of radiation to ICIs incrementally improves the systemic response to ICIs. Neoadjuvant therapy of ICIs plus stereotactic body radiotherapy (SBRT) has shown promising results in several types of solid tumours but not HCC. METHODS AND ANALYSIS: Here, we describe a phase Ib clinical trial of neoadjuvant SBRT plus PD-1 (tislelizumab) prior to hepatic resection in HCC patients. Prior to resection, eligible HCC patients will receive 8 Gy×3 fractions of SBRT together with two cycles of tislelizumab with an interval of 3 weeks. HCC resection is scheduled 4 weeks after the second dose of tislelizumab, followed by adjuvant tislelizumab for 1 year. We plan to enrol 20 participants in this trial. The primary study endpoints include the delay of surgery, tumour response and safety and tolerability of the sequential SBRT/tislelizumab. Other endpoints are the disease-free survival and overall survival rates every 3 or 6 months after the surgery. ETHICS AND DISSEMINATION: This trial was approved by the Ethics Committee of Shandong Cancer Hospital and Institute (SDZLEC2022-021-01). The final results of this trial will be published in a peer-reviewed journal after completion. TRIAL REGISTRATION NUMBER: NCT05185531.