Identification of immune-associated biomarker for predicting lung adenocarcinoma: bioinformatics analysis and experiment verification of PTK6.

BACKGROUND: Abnormal expression of protein tyrosine kinase 6 (PTK6) has been proven to be involved in the development of gynecological tumors. However, its immune-related carcinogenic mechanism in other tumors remains unclear. OBJECTIVE: The aim of this study was to identify PTK6 as a novel prognostic biomarker in pan-cancer, especially in lung adenocarcinoma (LUAD), which is correlated with immune infiltration, and to clarify its clinicopathological and prognostic significance. METHODS: The prognostic value and immune relevance of PTK6 were investigated by using bio-informatics in this study. PTK6 expression was validated in vitro experiments (lung cancer cell lines PC9, NCI-H1975, and HCC827; human normal lung epithelial cells BEAS-2B). Western blot (WB) revealed the PTK6 protein expression in lung cancer cell lines. PTK6 expression was inhibited by Tilfrinib. Colony formation and the Cell Counting Kit-8 (CCK-8) assay were used to detect cell proliferation. The wound healing and trans-well were performed to analyze the cell migration capacity. Then flow cytometry was conducted to evaluate the cell apoptosis. Eventually, the relationship between PTK6 and immune checkpoints was examined. WB was used to estimate the PD-L1 expression at different Tilfrinib doses. RESULTS: PTK6 was an independent predictive factor for LUAD and was substantially expressed in LUAD. Pathological stage was significantly correlated with increased PTK6 expression. In accordance with survival analysis, poor survival rate in LUAD was associated with a high expression level of PTK6. Functional enrichment of the cell cycle and TGF-ß signaling pathway was demonstrated by KEGG and GSEA analysis. Moreover, PTK6 expression considerably associated with immune infiltration in LUAD, as determined by immune analysis. Thus, the result of vitro experiments indicated that cell proliferation and migration were inhibited by the elimination of PTK6. Additionally, PTK6 suppression induced cell apoptosis. Obviously, PD-L1 protein expression level up-regulated while PTK6 was suppressed. CONCLUSION: PTK6 has predictive value for LUAD prognosis, and could up regulated PD-L1.

Attitudes toward communication skills with learner needs assessment within radiology residency programs in China: a cross-sectional survey.

BACKGROUND: Communication skills (CS) represent a core competency in radiology residency training. However, no structured curriculum exists to train radiology residents in CS in China. The aim of this study was to evaluate the status and prevalence of doctor-patient communication training among radiology residents in nine Chinese accredited radiology residency training programs and to determine whether there is a perceived need for a formalized curriculum in this field. METHODS: We administered a cross-sectional online survey to radiology residents involved in CS training at nine standard residency training programs in China. The questionnaire developed for this study included CS training status, residents' demographics, attitudes toward CS training, communication needs, and barriers. Residents' attitudes toward CS training were measured with the Communication Skills Attitude Scale (CSAS) and its subscales, a positive attitude scale (PAS) and negative attitude scale (NAS). RESULTS: A total of 133 (48.36%) residents participated in the survey. The mean total scores on the two dimensions of the CSAS were 47.61 ± 9.35 in the PAS and 36.34 ± 7.75 in the NAS. Factors found to be significantly associated with the PAS included receiving previous training in CS, medical ethics, or humanities and the doctor's attire. We found that first-year residents and poor personal CS were the most influential factors on the NAS. Only 58.65% of participants reported having previously received CS training during medical school, and 72.93% of respondents reported failure in at least one difficult communication during their residency rotation. Most of those surveyed agreed that CS can be learned through courses and were interested in CS training. Some of the most common barriers to implementing formal CS training were a lack of time, no standardized curriculum, and a lack of materials and faculty expertise. CONCLUSIONS: Most residents had a very positive attitude toward CS training and would value further training, despite the limited formal CS training for radiology residents in China. Future efforts should be made to establish and promote a standard and targeted CS curriculum for Chinese radiology residents.

HNRNPA2B1 Demonstrates Diagnostic and Prognostic Values Based on Pan-Cancer Analyses.

Some studies have suggested heterogeneous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1) to be a promoter in cancer development. Nonetheless, no detailed pan-cancer investigation has been reported. Thus, this study explored the possible oncogenic role of HNRNPA2B1, such as its expression levels, gene alteration, protein-protein interaction network, immune infiltration, and prognostic value in different cancer types using The Cancer Genome Atlas web platform. Many types of cancer exhibit HNRNPA2B1 overexpression, which is notably associated with poor prognosis. We also found that HNRNPA2B1 with different methylation levels causes a varied prognosis in lung adenocarcinoma (LUAD). It is noteworthy that HNRNPA2B1 levels are connected with cancer-associated fibroblasts in cancers, such as adrenocortical carcinoma, LUAD, and stomach adenocarcinoma. In addition, HNRNPA2B1 participates in the spliceosome- and cell cycle-associated pathways. Finally, HNRNPA2B1 is highly valued in the diagnosis of LUAD, lung squamous cell carcinoma, breast invasive carcinoma, esophageal carcinoma, and liver hepatocellular carcinoma. This systematic study highlighted the role of HNRNPA2B1 in pan-cancer progression.

Long noncoding RNA NR2F1-AS1 stimulates the tumorigenic behavior of non-small cell lung cancer cells by sponging miR-363-3p to increase SOX4.

Long noncoding RNA (lncRNA), specifically the upregulation of lncRNA NR2F1 antisense RNA 1 (NR2F1-AS1), has been involved in the progression of non-small cell lung cancer (NSCLC), but the mechanisms that underlie this remain unclear. In this study, the expression of NR2F1-AS1, miR-363-3p, and SOX4 was assessed in NSCLC cells. A loss-of-function assay was used to measure the tumorigenicity of NSCLC cells. The glycolysis and glutamine metabolism of NSCLC cells was also measured via extracellular acidification rate, consumption of glucose and glutamine, and production of lactate and ATP. The relationships among NR2F1-AS1, miR-363-3p, and SOX4 were detected via dual-luciferase reporter assay. HK-2, GLS1, and SOX4 levels were also analyzed. We found that both NSCLC tissues and cells had higher levels of NR2F1-AS1. Silencing of NR2F1-AS1 inhibited the tumorigenicity of cells in vitro and reduced the glycolysis and glutamine metabolism of NSCLC cells. Regarding its mechanism, NR2F1-AS1 positively regulated the SOX4 level by sponging miR-363-3p. Furthermore, miR-363-3p inhibition or SOX4 overexpression reversed the repressing role of sh-NR2F1-AS1 in the tumorigenicity of NSCLC cells. In summary, NR2F1-AS1 promotes the tumorigenicity of NSCLC cells by regulating miR-363-3p/SOX4.

Sulforaphane protects against oxidative stress­induced apoptosis via activating SIRT1 in mouse osteoarthritis.

Osteoarthritis (OA), the most common form of human joint disease, is characterized by progressive degeneration of the articular cartilage, synovitis and subchondral osteoporosis. Chondrocyte apoptosis is the primary pathogenic mechanism of OA and is considered to be a potential therapeutic target. Sulforaphane (SFN), a dietary isothiocyanate obtained from cruciferous vegetables, has been reported to exert an anti­apoptotic effect by activating sirtuin 1 (SIRT1). To the best of our knowledge, however, the effects of SFN on apoptotic responses in OA have not been reported. In the present study, SFN was shown to significantly inhibit chondrocyte apoptosis while enhancing expression levels of SIRT1 in a H2O2­induced OA mouse model. The anti­apoptotic effect of SFN was reversed by SIRT1 small interfering RNA, implying that SIRT1 exerted a protective role against the effect of SFN on chondrocytes. The expression levels of C/EBP homologous protein, 78­kDa glucose regulated protein, Bax, Bcl­2 and cleaved caspase 3 were found to be downregulated in SFN­treated mice. Furthermore, SFN ameliorated cartilage degradation in the OA mouse model. These findings indicate that SFN exerted an anti­apoptotic effect on chondrocytes and ameliorated OA in vivo by activating the SIRT1 signaling pathway.

[Pressure side and tension side comminution of femoral neck cortex are independent risk factors for aseptic necrosis after femoral neck fracture surgery].

OBJECTIVE: To investigate the related factors of aseptic necrosis of femoral head after closed reduction and internal fixation of femoral neck fracture. METHODS: From January 2009 to January 2016, 236 patients with femoral neck fracture were treated with closed reduction and internal fixation with 3 hollow lag screws, including 111 males and 125 females, aged from 19 to 89 (50.17±12.88) years. According to the follow-up results, the correlation of aseptic necrosis of femoral head was analyzed. Univariate analysis of age, gender, injured side, body weight, injury mechanism, preoperative waiting time, Garden classification and whether there was comminution of femoral neck cortex was conducted to obtain the independent variables with significant difference. Then binary logistic regression analysis was conducted to explore the independent risk factors of avascular necrosis of femoral head. RESULTS: The average follow-up period of 236 cases was 4.58 years. There were significant differences in the range of injury (24.69% vs. 5.16%, χ2=19.405, P=0.000), operation waiting time>48 hours (20.00% vs. 6.38%, χ2=10.065, P=0.002), Garden type Ⅲ/Ⅳ (18.52% vs. 2.97%, χ2=13.357, P=0.000), femoral neck cortex comminution (66.67% vs. 4.88%, χ2=39.968, P=0.000). Multivariate logistic regression analysis showed that:injury mechanism [high energy injury, Exp (B)=4.397, 95%CI=(1.672-11.562), P=0.003], preoperative waiting time >48 h [Exp (B)= 3.060, 95%CI=(1.176-7.966), P=0.022], comminution of femoral neck cortex [comminution of femoral neck pressure side cortex, Exp (B)=3.944, 95%CI=(1.245-12.494), P=0.020;comminution of femoral neck pressure side and tension side cortex, Exp(B)= 23.761, 95%CI=(3.805-148.374), P=0.001) were independent risk factors for avascular necrosis after internal fixation of femoral neck fracture. Garden type Ⅲ/Ⅳ was not an independent risk factor in this study [Exp (B) = 1.985, 95%CI=(0.436-9.032), P=0.375]. CONCLUSION: High energy injury, preoperative waiting time (>48 h) and comminution of femoral neck cortex were independent risk factors for aseptic necrosis of femoral head. In addition, cortical comminution on the pressure side and tension side of the femoral neck is a strong prognostic risk factor for aseptic necrosis of the femoral head, because it indicates a more serious and complex injury mechanism.

Salvianolic acid A blocks vasculogenic mimicry formation in human non-small cell lung cancer via PI3K/Akt/mTOR signalling.

Vasculogenic mimicry (VM) is associated with aggressive cancer cells. Salvianolic acid A (Sal-A), an antioxidant and anti-inflammatory agent, has bioactive properties from Salvia miltiorrhiza Bunge. Current investigation aspired to explore the activity of Sal-A in the VM formation of non-small cell lung cancer (NSCLC) and the mechanism underling this function. The CCK8, the scratch and boyden chemotaxis assay were presented to describe NSCLC cells viability, migration and invasion capabilities, respectively. The protein expression was verified by western blotting. In this report, Sal-A caused a reduction in viability, metastasis and capillaries structure formation of NSCLC cells. Additionally, Sal-A markedly prevented the key VM related proteins, containing EphA2, VE-cadherin and MMP2. Besides, Sal-A significantly diminished p-PI3K, p-Akt and p-mTOR level in NSCLC cells. More importantly, SC79 pretreatment reversed Sal-A inhibits NSCLC cells viability, metastasis and VM formation. These data exhibit that Sal-A could block VM network formation in NSCLC cells through modulating the PI3K/Akt/mTOR signalling pathway.

Retraction notice to "The influence of connective tissue growth factor on rabbit ligament injury repair" [Saudi Pharm. J. 25 (2017) 498-503].

[This retracts the article DOI: 10.1016/j.jsps.2017.04.013.].

Nobiletin Inhibits IL-1ß-Induced Inflammation in Chondrocytes via Suppression of NF-κB Signaling and Attenuates Osteoarthritis in Mice.

Osteoarthritis (OA), a common degenerative joint disease, is principally characterized by inflammation and destruction of cartilage. Nobiletin, an extract of the peel of citrus fruits, is known to have anti-inflammatory properties. However, the mechanisms by which nobiletin plays a protective role in osteoarthritis (OA) are not completely understood. In the present study, we investigated the anti-inflammatory effects of nobiletin in the progression of OA in both in vitro and in vivo experiments. Mouse chondrocytes were pretreated with nobiletin (0, 10, 20, 40 µM) for 24 h and then incubated with IL-1ß (10 ng/ml, 24 h) in vitro. The generation of PGE2 and NO was evaluated by the Griess reaction and ELISAs. The protein expression of inducible nitric oxide synthase, matrix metalloproteinase-3, matrix metalloproteinase-13, A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS5), cyclooxygenase-2, collagen II, and aggrecan was analyzed by Western blotting. Immunofluorescence and Western blot analysis were used to detect nuclear factor-κB (NF-κB) signaling molecules. Induction of proinflammatory and catabolic mediators by IL-1ß stimulation of mouse chondrocytes could be partially blocked by treatment with nobiletin or ammonium pyrrolidine dithiocarbamate (an NF-κB inhibitor). Furthermore, our results indicated that nobiletin exhibited a therapeutic effect through active inhibition of the NF-κB signaling pathway. In a mouse model of OA, injection of nobiletin (20 mg/kg) every 2 days for 8 weeks after surgery inhibited cartilage destruction and synovitis. Taken together, our findings suggest that nobiletin may be a potential therapeutic agent for the treatment of OA.

Human Mind Control of Rat Cyborg's Continuous Locomotion with Wireless Brain-to-Brain Interface.

Brain-machine interfaces (BMIs) provide a promising information channel between the biological brain and external devices and are applied in building brain-to-device control. Prior studies have explored the feasibility of establishing a brain-brain interface (BBI) across various brains via the combination of BMIs. However, using BBI to realize the efficient multidegree control of a living creature, such as a rat, to complete a navigation task in a complex environment has yet to be shown. In this study, we developed a BBI from the human brain to a rat implanted with microelectrodes (i.e., rat cyborg), which integrated electroencephalogram-based motor imagery and brain stimulation to realize human mind control of the rat's continuous locomotion. Control instructions were transferred from continuous motor imagery decoding results with the proposed control models and were wirelessly sent to the rat cyborg through brain micro-electrical stimulation. The results showed that rat cyborgs could be smoothly and successfully navigated by the human mind to complete a navigation task in a complex maze. Our experiments indicated that the cooperation through transmitting multidimensional information between two brains by computer-assisted BBI is promising.

Circular RNA circNASP modulates the malignant behaviors in osteosarcoma via miR-1253/FOXF1 pathway.

Increasing evidences demonstrate that circular RNAs (circRNAs) serve as essential regulators in various human diseases, including cancer. However, the study on circRNA function in osteosarcoma (OS) is just emerging. In the present study, we screened out a novel circRNA termed circNASP which was significantly upregulated in OS tissues compared to adjacent normal tissues. We found that circNASP knockdown dramatically inhibited the proliferation, cell cycle progression and invasion of OS cells. Moreover, we showed that circNASP expression was positively correlated with tumor size and metastasis in OS patients. In terms of mechanism, we found that circNASP acts as a sponge of miR-1253 targeting FOXF1 in OS cells. By inhibiting miR-1253 availability, circNASP promoted FOXF1 expression. Rescue assays indicated that inhibition of miR-1253 could significantly reverse the effects of circNASP knockdown on OS cell proliferation and invasion while restoration of FOXF1 rescued the proliferation and invasion of OS cells transfected with miR-1253 mimics. Taken together, our findings demonstrated that circNASP contributes to malignant behaviors of OS cells by miR-1253/FOXF1 pathway, which suggested circNASP/miR-1253/FOXF1axis might be a potential therapeutic target.

The Effects of Aqueous Extract from Nardostachys chinensis Batalin on Blood Pressure and Cardiac Hypertrophy in Two-Kidney One-Clip Hypertensive Rats.

AIMS: The aim of this study was to investigate the effects of the aqueous extract of Nardostachys chinensis Batalin (NCBAE) on blood pressure and cardiac hypertrophy using two-kidney one-clip (2K1C) hypertensive rats. METHODS: 2K1C rat models were set up by clipping the left renal artery. Sham-operated rats underwent the same surgical procedure except for renal arterial clipping. 2K1C hypertensive rats were orally given NCBAE at doses of 210, 420, and 630 mg·kg-1·d-1 for 6 weeks. Twelve weeks after surgery, rat SBP and echocardiographic parameters were measured, cardiac histopathology was assessed, serum NO and LDH were detected, and the expression of Bcl-2 and caspase-3 of left ventricular tissue was assessed by western blot. RESULTS: Treatment with NCBAE resulted in a decrease of SBP, LVPWd, LVPWs, IVSd, IVSs, LVW/BW ratio, and cardiomyocyte CSA, an increase of LVEF, and inhibition of 2K1C-induced reduction in serum NO and elevation of LDH compared with 2K1C group. NCBAE intervention also showed a significant increase of Bcl-2 expression and reduction of cleaved caspase-3 level dose-dependently in left ventricular tissue. CONCLUSION: Our data demonstrate that NCBAE has an antihypertensive property and protective effect on 2K1C-induced cardiac hypertrophy especially at the dose of 630 mg·kg-1·d-1.

The influence of connective tissue growth factor on rabbit ligament injury repair.

OBJECTIVES: The injured anterior cruciate ligament (ACL) is deemed to exhibit an impaired healing response and attempts at surgical repair have not been successful. Connective tissue growth factor (CTGF) is reported to be associated with wound healing, probably through transforming growth factor beta1 (TGF-ß1). METHODS: A rabbit ACL injury model was used to study the effect of CTGF on ligament recovery. Quantitative real-time PCR was performed for detection of changes in RNA levels of TGF-ß1, type 1 collagen (COL-I), type 2 collagen (COL-II), SRY-related high mobility group-box gene9 (Sox9), metalloproteinase-1 (TIMP-1) as well as matrix metallopeptidase 13 (MMP-13). And expression of related proteins was detected by western blotting. RESULTS: The current study showed that CTGF could promote the recovery of inured anterior cruciate ligament. It can up-regulate the mRNA and expression of TGF-ß1, COL-I, COL-II, Sox9, as well as the tissue inhibitor of TIMP-1, and down-regulated the mRNA and expression of MMP-13, suggesting the curative effect of CTGF on injured rabbit ligament is through regulating these cellular factors. CONCLUSION: This finding revealed the mechanism of CTFG's healing role in injured tissues and provided new possibilities of treating injured tissues and wound healing by using CTFG.

BEM-based simulation of lung respiratory deformation for CT-guided biopsy.

PURPOSE: Accurate and real-time prediction of the lung and lung tumor deformation during respiration are important considerations when performing a peripheral biopsy procedure. However, most existing work focused on offline whole lung simulation using 4D image data, which is not applicable in real-time image-guided biopsy with limited image resources. In this paper, we propose a patient-specific biomechanical model based on the boundary element method (BEM) computed from CT images to estimate the respiration motion of local target lesion region, vessel tree and lung surface for the real-time biopsy guidance. METHODS: This approach applies pre-computation of various BEM parameters to facilitate the requirement for real-time lung motion simulation. The resulting boundary condition at end inspiratory phase is obtained using a nonparametric discrete registration with convex optimization, and the simulation of the internal tissue is achieved by applying a tetrahedron-based interpolation method depend on expert-determined feature points on the vessel tree model. A reference needle is tracked to update the simulated lung motion during biopsy guidance. RESULTS: We evaluate the model by applying it for respiratory motion estimations of ten patients. The average symmetric surface distance (ASSD) and the mean target registration error (TRE) are employed to evaluate the proposed model. Results reveal that it is possible to predict the lung motion with ASSD of [Formula: see text] mm and a mean TRE of [Formula: see text] mm at largest over the entire respiratory cycle. In the CT-/electromagnetic-guided biopsy experiment, the whole process was assisted by our BEM model and final puncture errors in two studies were 3.1 and 2.0 mm, respectively. CONCLUSION: The experiment results reveal that both the accuracy of simulation and real-time performance meet the demands of clinical biopsy guidance.

[The causes and preventive methods of misdiagnosis on wrist scaphoid bone fracture].

OBJECTIVE: To retrospectively analysis the curative effect of wrist scaphoid bone fracture,and explore the causes and preventive methods of misdiagnosis. METHODS: From September 2007 to September 2010,16 patients with wrist scaphoid bone fractures were treated with plaster cast and cannulated screws fixation. There were 10 males and 6 females,ranging in age from 26 to 44 years with an average of 35 years. Among them, 12 cases manifested swelling pain of radial lateral wrist, tenderness at snuffbox area, wrist pain aggravated when stretching wrist joint, thumb or forefinger; 4 cases manifested no obviously symptoms and limited movement; 9 cases were early diagnosed; 5 cases were treated by plaster cast; 4 cases were treated with cannulated screws fixation; Among 7 cases with misdiagnosis, there were 4 cases without obvious symptoms and they were dealt with activating blood to dissipate swelling and pain process in preliminary stage. Four cases were treated with plaster cast and 3 cases with cannulated screws fixation. RESULTS: All the patients were followed up from 3 months to 39 months (averaged 21 months). Among 16 patients, 9 cases were early diagnosis, 7 cases were misdiagnosis and the rate of misdiagnosis was 43.8%. Seven cases with screws fixation were no wound infection. There was 1 case with occurred chronic pain and declining wrist mobility in both plaster cast and screw group, and both of them were misdiagnosed. According to curative effect rating criteria,these 2 cases were classified into moderate, other 14 cases were excellent. CONCLUSION: Wrist scaphoid bone fracture are easy to misdiagnose, so early diagnosis and treatment is particularly important. The main causes of misdiagnosis are nonspecific symptoms at early stage, combination with other injuries, lack of knowledge and ignorance of the further examination. Therefore, detailed inquiries and particular examination, multi-dimensional radiography and CT scan or MRI scan are the main measures for prevention.

[Application of endobutton in the treatment of acute acromioclavicular joint dislocation].

OBJECTIVE: To study the clinical efficacy of the endobutton in the treatment of acute acromioclavicular joint dislocation by reconstructing coracoclavicular ligaments. METHODS: From October 2008 to January 2010,12 patients with acute acromioclavicular joint dislocation were immobilized with the endobutton. All the patients had the dislocations of or above type III according to Rockwood classification. Among the patients, 9 patients were male and 3 patients were female, with an average age of 55 years (ranged from 31 to 83 years). Eight patients had injuries in the left, and 4 patients in the right. Four patients had accompanied injuries of rib fractures, 2 patients had brain injuries,and 1 patient had femoral fracture. Seven patients were injured by traffic accident, 4 patients were injured by falling down,and 1 patient was sports injuries. All the patients had pain and tenderness at the shoulder, positive piano sign, and shoulder confined activity. The duration from injury to operation ranged from 2 days to 10 days (averaged 6 days). The therapeutic effects were evaluated by Karlsson criteria based on range of motion of acromioclavicular joint, subjective feeling,and postoperative X-ray. RESULTS: All the patients were followed up, and the duration ranged from 4 months to 19 months (averaged 11 months). The motion of the shoulder joint recovered to normal about 15 to 35 days after operation. There were no displacement, dislocation and redislocation occurred. All the patients got A degree results according to Karlsson criteria. CONCLUSION: Reconstruction of coracoclavicular ligament by using the endobutton to treat acute acromioclavicular dislocation of or above type III is a perfect method with advantage of rigid fixation, micro-injury, and early functional exercise.

[Impact of radiofrequency on splenocyte immunity of mice bearing H22 liver cancer].

BACKGROUND & OBJECTIVE: Radiofrequency (RF) can destroy local tumor tissue with less damage to the intervening tissue. The effect of RF on immune function of tumor-bearing body needs to be further investigated. This study was designed to evaluate the impact of RF on splenocyte immunity of mice bearing H22 liver cancer through detecting splenocyte proliferation, cytotoxicity, and expression pattern of cytokines T helper type 1 (Th1)/Th2. METHODS: A total of 24 BALB/c mice were randomized into 4 groups: RF, surgical resection, tumor-bearing control, and normal control groups. The proliferation of splenocytes was detected by MTT assay, and the cytotoxicity of splenocytes was assayed by double-color flow cytometry (FCM). The concentrations of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-4, and IL-10 in splenocyte culture supernatants were detected by enzyme-linked immunoabsorbent assay (ELISA), and mRNA levels of the cytokines in splenocytes were assayed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The proliferation of splenocytes and mortality of H22 cells were significantly higher in RF group than in surgical resection group, normal control group, and tumor-bearing group (P<0.05). The concentration and mRNA level of IL-2 and IFN-gamma were significantly higher in RF group than in surgical resection group, normal control group, and tumor-bearing group (P<0.05); the concentration and mRNA level of IL-4 and IL-10 were significantly lower in RF group than in surgical resection group, normal control group, and tumor-bearing group (P<0.05). No significant difference were found between surgical resection group and normal control group (P>0.05). CONCLUSIONS: RF can efficiently stimulate splenocyte activation and proliferation in H22-bearing mice, and enhance splenocyte cytotoxicity to tumor cells. RF could facilitate the secretion and gene transcription of Th1 type cytokines in H22 cells.