Primary tumour location is an important prognostic factor in colorectal cancer liver metastases following radiofrequency ablation.

AIM: To examine the outcomes and different prognoses for colorectal cancer patients with liver metastases (CRCLM) treated with radiofrequency ablation (RFA) based on the origin of the primary tumour (rectum versus colon). MATERIALS AND METHODS: Patients diagnosed with CRCLM from July 2011 to June 2018 were assessed in the study. The study included 114 patients with 176 lesions. All of the patients underwent RFA of their CRCLM. The overall survival (OS) and progression-free survival (PFS) were assessed with Kaplan-Meier survival function estimate curves and Cox regression analysis between the group of patients with rectal cancer liver metastases (RCLM) and colon cancer liver metastases (CCLM). RESULTS: Patients with RCLM exhibited worse OS and PFS than those with CCLM. The 1-, 2-, and 3-year OS rates were 87.5%, 75%, and 54.2% in the RCLM group and 93.9%, 87.8%, and 81.8% in the CCLM group (p=0.001), respectively. The 1-, 2-, and 3-year PFS rates were 43.7%, 22.9%, and 20.8% in the RCLM group and 57.6%, 45.4%, and 36.5% in the CCLM group (p=0.001), respectively. Multivariate analysis showed that primary tumour location, carcinoembryonic antigen (CEA), and response to pre-ablation chemotherapy were independent variables for OS and PFS. The N state of the primary tumour also had a significant impact on OS. CONCLUSION: This study showed that patients with CCLM had better prognoses than those with RCLM following RFA treatment. If patients intend to undergo RFA for CRCLM, the location of the primary tumour should be considered in the assessment of long-term therapeutic effects.

Altered mechanics of vaginal smooth muscle cells due to the lysyl oxidase-like1 knockout.

The remodeling mechanisms that cause connective tissue of the vaginal wall, consisting mostly of smooth muscle, to weaken after vaginal delivery are not fully understood. Abnormal remodeling after delivery can contribute to development of pelvic organ prolapse and other pelvic floor disorders. The present study used vaginal smooth muscle cells (vSMCs) isolated from knockout mice lacking the expression of the lysyl oxidase-like1 (LOXL1) enzyme, a well-characterized animal model for pelvic organ prolapse. We tested if vaginal smooth muscle cells from LOXL1 knockout mice have altered mechanics including stiffness and surface adhesion. Using atomic force microscopy, we performed nanoindentations on both isolated and confluent cells to evaluate the effect of LOXL1 knockout on in vitro cultures of vSMCs cells from nulliparous mice. The results show that LOXL1 knockout vSMCs have increased stiffness in pre-confluent but decreased stiffness in confluent cultures (p* < 0.05) and significant decreased surface adhesion in pre-confluent cultures (p* < 0.05). This study provides evidence that the weakening of vaginal connective tissue in the absense of LOXL1 changes the mechanical properties of the vSMCs. STATEMENT OF SIGNIFICANCE: Pelvic organ prolapse is a common condition affecting millions of women worldwide, which significantly impacts their quality of life. Alterations in vaginal and pelvic floor mechanical properties can change their ability to support the pelvic organs. This study provides evidence of altered stiffness of vaginal smooth muscle cells from mice resembling pelvic organ prolapse. The results from this study set a foundation to develop pathophysiology-driven therapies focused on the interplay between smooth muscle mechanics and extracellular matrix remodeling.

Genomic and transcriptional heterogeneity of multifocal hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) often presents with multiple nodules within the liver, with limited effective interventions. The high genetic heterogeneity of HCC might be the major cause of treatment failure. We aimed to characterize genomic heterogeneity, infer clonal evolution, investigate RNA expression pattern and explore tumour immune microenvironment profile of multifocal HCC. PATIENTS AND METHODS: Whole-exome sequencing and RNA sequencing were carried out in 34 tumours and 6 adjacent normal liver tissue samples from 6 multifocal HCC patients. Protein expression of Ki67, AFP, P53, Survivin and CD8 was detected by immunohistochemistry. Fluorescence in situ hybridization was carried out to validate the amplification status of sorafenib-targeted genes. RESULTS: We deciphered genomic and transcriptional heterogeneity among tumours in each multifocal HCC patient including mutational profiles, copy number alterations, tumour evolutionary trajectory and tumour immune microenvironment profiles. Of note, sorafenib-targeted alterations were identified in the trunk of phylogenetic tree in only one out of the six patients, which may explain the relative low treatment response rate to sorafenib in clinical practice. Moreover, we demonstrated RNA expression patterns and tumour immune microenvironment profiles of all nodules. We found that RNA expression pattern was associated with Edmondson-Steiner grading. Based on the differential expression of 66 reported immune markers, unsupervised hierarchical clustering analysis of 34 nodules identified immune subsets: one low expression cluster with seven nodules and one high expression cluster with 11 nodules. CD8+ T cells were more enriched in nodules of the high expression cluster. CONCLUSIONS: Our study provided a detailed view of genomic and transcriptional heterogeneity, clonal evolution and immune infiltration of multifocal HCC. The heterogeneity of druggable targets and immune landscape might help interpret the clinical responsiveness to targeted drugs and immunotherapy for multifocal HCC patients.

[Establishment and evaluation of a new method for determining hemodynamics of pulmonary hypertension rats].

Objective: By evaluating the hemodynamic parameters such as cardiac output (CO), right ventricular pressure (RVP), pulmonary artery pressure (PAP) and total pulmonary resistance index (TPRI) in pulmonary hypertension rat model, we established a more comprehensive hemodynamic evaluation system, which objectively evaluated the severity of disease and exercise tolerance in rats with pulmonary hypertension. Methods: SD rats were randomly divided into a control group and a model group with 5 rats in each group. The model group was intraperitoneally injected with SU5416 (20 mg/kg) and placed in an oxygen chamber at a 10% oxygen concentration for 21 days and then placed in a normoxic environment for 14 days. After modeling, rats were anesthetized and mechanically ventilated. The operator cut the skin along the right paraxial line, detached and ligated the intercostal artery, and then cut off the 3 and 4 ribs, exposing the heart and freeing aortic root about 0.2 cm. The flowmeter probe was set in the dissected aortic segment, and real-time recording time, blood flow waveforms, cardiac output were calculated accordingly. Then the needle attached to the baroreceptor was inserted into the right ventricle and the system acquired the right ventricular time-pressure waveform. After the waveform stabilized for about 30 seconds, the end of the cannula was sent to the pulmonary artery trunk through the entrance of the pulmonary artery to record the time-pressure curve of the pulmonary artery. Results: RVSP, PASP, PADP and mPAP in the model group were significantly higher than those of the control group [ RVSP(23.4±5.4) mmHg, 1 mmHg=0.133 kPa vs (56.4±13.0) mmHg, PASP (22.8±4.4) mmHg vs (58.5±14.9) mmHg, PADP (9.7±1.9) mmHg vs (30.3±7.0) mmHg, mPAP (14.1±2.7) mmHg vs (41.9±8.0) mmHg, all P<0.05 ]. Compared with the control group, the cardiac index in the model group was significantly lower [ CI (0.54±0.08) ml·min(-1)·g(-1) vs (0.40±0.09) ml·min(-1)·g(-1,) P=0.02 ]. Furthermore, compared with the control group, pulmonary vascular resistance index was significantly increased in the model group[PVRI (0.27±0.03) mmHg·ml(-1)·min(-1)·kg(-1) vs (0.06±0.01) mmHg·ml(-1)·min(-1)·kg(-1,) P<0.05]. The pathological results also showed that the middle part of pulmonary arterioles in the model group had muscular hypertrophy and muscular pulmonary arterioles, and even plexiform lesions. Conclusion: In this study, we established a new method that simultaneously determined several hemodynamic parameters such as RVSP, PASP, PADP, CO, CI and PVRI, which provided a more comprehensive assessment of hemodynamic changes in pulmonary hypertension rat models.

Treatment of hepatocellular carcinoma in the caudate lobe: US-guided percutaneous radiofrequency ablation combined with ethanol ablation.

AIM: To evaluate the technical feasibility and treatment results of ultrasound (US)-guided percutaneous radiofrequency ablation (RFA) combined with ethanol ablation (EA) for hepatocellular carcinoma (HCC) in the caudate lobe. MATERIALS AND METHODS: The institutional review board approved this retrospective study and the requirement for informed consent was waived. From October 2009 to June 2016, there were 49 patients with HCC in the caudate lobe underwent ablation. Among them, 14 patients (12 men and two women, mean age: 49.9±14.9 years) with 16 caudate lobe HCCs (mean size: 3±1.1 cm) who underwent US-guided RFA-EA were identified. These patients were compared with patients who underwent RFA alone (n=17) and EA alone (n=18). RESULTS: HCCs in the RFA-EA group were larger than those in the EA-alone group (3±1.1 versus 2.1±0.7 cm, p=0.028). In the RFA-EA group, all the patients achieved treatment success (14/14) and entered follow-up. After a follow-up period of 17.1±10.6 months (range, 5-36 months), local tumour progression (LTP) was detected in four tumours. The 1- and 2-year LTP rates after RFA-EA were 18.8% and 27.8%, respectively. The 1- and 2-year disease-free survival (DFS) rates were 35.7% and 13.4%. Six patients died, with the 1-, 2-, and 3-year overall survival (OS) rates of 85.1%, 63.1%, and 21%, respectively. There were no significant differences in the LTP, DFS, and OS rates between the RFA-EA group and RFA-alone or EA groups (p=0.363-0.733). CONCLUSION: US-guided percutaneous RFA-EA appears to be a feasible and effective treatment for HCC in the caudate lobe.

[Method for recording tension changes of small pulmonary artery and vein using the isolated vessel tension measurement system].

Objective: To establish a standardized method for isolated pulmonary artery and vein rings with different diameter, pressure and length, which could provide a more scientific method for in vitro study of pulmonary vessel diseases. Methods: Male SD rats were anesthetized, and the right ventricular systolic pressure were measured. Small pulmonary artery and vein rings with 200-400 µm in diameter and 2 mm in length were prepared by dissecting pulmonary arteries and veins. The pulmonary vessel rings were mounted in the organ bath by 2 stainless steel wires with diameter of 40 µm. Then the internal circumference of the vessel rings was increased gradually with 100 µm per step. At the same time the vascular tension was recorded by the Myograph System and Acknowledgement data acquisition system, and subsequently the passive length-tension exponential curve was made. The initial tension of the rings was set, equilibrated for another 30 min, and then stimulated with 60 mmol/L KCl 3 times, and the best contractile reactivity was achieved. The contractile reactivity of pulmonary artery rings and endothelial integrity were detected by exposure to 1 µmol/L phenylephrine(PE) and 10 µmol/L acetylcholine(Ach), while the contractile reactivity of pulmonary vein rings was detected by exposure to 1 µmol/L U46619 and 10 µmol/L papaverine. Results: The contraction and relaxation effects of the pulmonary artery rings reached 0.39 mg and 92% when they were stimulated by 1 µmol/L PE and 10 µmol/L Ach. The contraction and relaxation effects of pulmonary vein rings were up to 0.13 mg and 84% when they were exposed to 1 µmol/L U46619 and 10 µmol/L papaverine, respectively. Conclusion: Pulmonary artery and vein rings with appropriate basal tension and optimal vasodilator activity were prepared, and a standardized method of tension experiment for isolated pulmonary artery and vein rings established.

Arterial spin labelling MRI for detecting pseudocapsule defects and predicting renal capsule invasion in renal cell carcinoma.

AIM: To evaluate prospectively the performance of combining morphological and arterial spin labelling (ASL) magnetic resonance imaging (MRI) for detecting pseudocapsule defects in renal cell carcinoma (RCC), and to predict renal capsule invasion confirmed histopathologically. MATERIALS AND METHODS: Twenty consecutive patients with suspicious renal tumours underwent MRI. Renal ASL imaging was performed and renal blood flow was measured quantitatively. The diagnostic performance of T2-weighted images alone, and a combination of T2-weighted and ASL images for predicting renal capsule invasion were assessed. RESULTS: Twenty renal lesions were evaluated in 20 patients. All lesions were clear cell RCCs (ccRCCs) confirmed at post-surgical histopathology. Fifteen ccRCCs showed pseudocapsule defects on T2-weighted images, of which 12 cases showed existing blood flow in defect areas on perfusion images. To predict renal capsule invasion, the sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 71.4%, 86.7%, 100%, respectively, for T2-weighted images alone, and 92.3%, 100%, 100%, 87.5%, respectively, for the combination of T2-weighted and ASL images. CONCLUSION: ASL images can reflect the perfusion of pseudocapsule defects and as such, the combination of T2-weighted and ASL images produces promising diagnostic accuracy for predicting renal capsule invasion.

Anatomical versus non-anatomical liver resection for hepatocellular carcinoma exceeding Milan criteria.

BACKGROUND: Liver resection is effective for hepatocellular carcinoma (HCC) exceeding the Milan criteria in selected patients. However, the benefit of anatomical resection (AR) versus non-anatomical resection (NAR) has not been clarified in this patient subgroup. This study aimed to compare outcomes between AR and NAR for HCC exceeding the Milan criteria. METHODS: Data on consecutive patients with HCC exceeding the Milan criteria who underwent liver resection with curative intent over a recent 6-year interval were extracted from a prospective single-centre HCC database and examined retrospectively. The postoperative outcomes of patients were compared before and after propensity score matching. RESULTS: Some 546 patients were included: 264 in the AR and 282 in the NAR group. In the original cohort, the AR group contained more patients with larger tumours, multiple tumours, macroscopic portal vein tumour thrombi, incomplete tumour capsules and microscopic vascular invasion. After propensity score matching, 177 pairs of patients were selected. The baseline data, including liver function and tumour burden, were similar in the matched groups. The 3-year recurrence-free survival rate was comparable between the matched NAR and AR groups (36·5 versus 28·5 per cent; P = 0·448). Similar results were observed for 3-year overall survival (57·5 versus 50·3 per cent; P = 0·385), recurrence patterns and early recurrence rates (57·6 per cent versus 59·9 per cent; P = 0·712). CONCLUSION: AR and NAR achieved favourable and similar outcomes for HCC exceeding the Milan criteria in selected patients.

Impact of Food Restriction on the Expression of the Adiponectin System and Genes in the Hypothalamic-Pituitary-Ovarian Axis of Pre-Pubertal Ewes.

Adiponectin, a cytokine secreted typically by adipocytes, has been implicated as a molecular switch between female reproduction and energy balance. The present study was undertaken to investigate the expression of adiponectin system and patterns of genes in the hypothalamic-pituitary-ovary (HPO) axis of food-restricted pre-pubertal ewes. Eighteen 2-month-old female ewes were assigned to 3 groups after a pre-feeding ad libitum for 10 days (six in each group): the control group (C), the low-food-restricted group (LR) and the high-food-restricted group (HR), which were fed with 100%, 70% and 50% of ad libitum food intake, respectively. The hypothalamus, pituitary, ovary and serum were collected after food restriction for 2 months. Results by ELISA showed that food restriction increased serum adiponectin concentrations. Quantitative real-time PCR showed that the gene transcriptions for adiponectin receptor 1 (AdipoR1) and 2 (AdipoR2) were enhanced in the hypothalamic-pituitary-ovarian (HPO) axis, while KISS-1/GPR-54 and gonadotropin-releasing hormone (GnRH) in the hypothalamus and luteinizing hormone ß-subunit (LHß) and follicle-stimulating hormone ß-subunit (FSHß) in the pituitary were reduced after food restriction. Immunohistochemistry results demonstrated that AdipoR1 localized in the oocytes of follicles in the ovary. These results suggest that the alterations in the expression of adiponectin and its receptors in response to food restriction might negatively influence the HPO axis.

Glutathione S-transferase P1 rs1695 A>G polymorphism and breast cancer risk: evidence from a meta-analysis.

Breast cancer (BC) is the most widespread cause of cancer-related deaths in women. Many published studies have assessed the association between the glutathione S-transferase P1 (GSTP1) rs1695 polymorphism and BC risk. However, the effect of the GSTP1 rs1695 polymorphism on BC risk has remained controversial. Therefore, this meta-analysis was conducted to obtain a comprehensive estimation of this association. A total of 20,615 cases and 20,481 controls from thirty-six case-control trials were extracted from an online literature survey. The meta-analysis indicated that the GSTP1 rs1695 A>G polymorphism did not contribute to the susceptibility of BC when the overall population was considered. However, intriguingly, this polymorphism was significantly associated with increased risk of BC in Asian women [GG vs AA: odds ratio (OR) = 1.4, 95% confidence interval (CI): 1.06-1.88, P = 0.02; AG vs AA: OR = 1.08, 95%CI = 1.00-1.16, P = 0.05; GG/AG vs AA: OR = 1.11, 95%CI = 1.04-1.19, P = 0.00]. Moreover, a subgroup analysis based on the source of control groups showed a marked increase in BC susceptibility in hospital-based control subjects (GG vs AA: OR = 1.28, 95%CI = 1.10-1.48, P= 0.00; GG vs AG/AA: OR = 1.22, 95%CI = 1.06-1.41, P = 0.00; GG/AG vs AA: OR = 1.10, 95%CI = 1.02-1.18, P = 0.00). In conclusion, our study indicated that the GSTP1 rs1695 A>G polymorphism was correlated with elevated BC risk in Asian women. Our results must be validated with further research.

Screening of highly informative and representative microsatellite markers for genotyping of major cultivated cotton varieties.

We screened and assessed published cotton simple sequence repeat (SSR) primers to establish a set of core SSR markers suitable for cotton major cultivars in China and analyzed genetic diversity based on the core marker set. Using a stepwise screening strategy, 12 leading cultivars for preliminary screening and 96 cultivars for rescreening were evaluated. A total of 184 polymorphic SSR markers were initially screened from 3299 candidates, and a core set of 52 SSR markers with wide genome coverage (2 markers per chromosome) was obtained. Among 96 major cultivars, 273 amplification genotypes were generated using the core marker set. Polymorphism information content values ranged from 0.28-0.83, with an average value of 0.56. The core SSR marker set detected on denaturing polyacrylamide gel electrophoresis indicated that the band genotype was either a single or double band on conventional cultivars, while most were double bands (65.4%). Among 56 hybrids, the average heterozygosis rate was 35.8%, ranging from 7.1-55.4%. Eighteen of 96 cultivars had distinct band genotypes. The genetic diversity analyzed using the of NTSYS-pc V2.10 software indicated that the Yangtze River valley cotton region had the highest polymorphic level, followed by Xinjiang and then the Yellow River valley. The genetic basis of conventional cultivars was narrower than that of hybrids. The core marker set can be used for fingerprint construction, variety identification, and purity tests of major cotton cultivars in China.

Construction of a primary DNA fingerprint database for cotton cultivars.

Forty core primers were used to construct a DNA fingerprint database of 132 cotton species based on multiplex fluorescence detection technology. A high first successful ratio of 99.04% was demonstrated with tetraplex polymerase chain reaction. Forty primer pairs amplified a total of 262 genotypes among 132 species, with an average of 6.55 per primer and values of polymorphism information content varying from 0.340 to 0.882. Conflicting DNA homozygous ratios were found in various species. The highest DNA homozygous ratio was found in landrace standard cultivars, which had an 81.46% DNA homozygous ratio. The lowest occurred in a group of 2010 leading cultivars with a homozygous ratio of 63.04%. Genetic diversity of the 132 species was briefly analyzed using unweighted pair-group method with arithmetic means.

Imaging features of hepatic angiomyolipomas on real-time contrast-enhanced ultrasound.

The aim of this study was to evaluate the imaging features of hepatic angiomyolipoma (AML) on contrast-enhanced ultrasound (CEUS). The imaging features of 12 pathologically proven hepatic AML lesions in 10 patients who had undergone baseline ultrasound (BUS) and CEUS examinations were evaluated retrospectively. The enhancement extent, pattern and dynamic change, along with the enhancement process, on CEUS were analysed. The diagnostic results of BUS and CEUS before pathological examination were also recorded. The results showed that 75% (9/12) of the AML lesions exhibited mixed echogenicity on BUS and most showed remarkable hyperechogenicity in combination with a hypoechoic or anechoic portion. Arterial flow signals were detected in 75% (9/12) of the lesions on colour Doppler imaging. On CEUS, 66.7% (n = 8) of the 12 lesions exhibited hyperenhancement in the arterial phase, slight hyperenhancement (n = 2) or isoenhancement (n = 6) in the portal phase, and slight hyperenhancement (n = 1) or isoenhancement (n = 7) in the late phase. Three (25%) lesions exhibited hyperenhancement in the arterial phase and hypoenhancement in both portal and late phases. One (8.3%) lesion exhibited hypoenhancement throughout the CEUS process. Before pathological examination with BUS, only 3 (25%) lesions were correctly diagnosed as hepatic AML. Conversely, on CEUS, correct diagnoses were made for 66.8% (8/12) of hepatic AMLs. Therefore, arterial hyperenhancement and subsequent sustained enhancement on CEUS were found in the majority of hepatic AMLs. The combination of BUS and CEUS leads to the correct diagnosis in the majority of hepatic AMLs, and is higher than the success rate achieved by BUS alone.

Prognostic factors for long-term outcome after percutaneous thermal ablation for hepatocellular carcinoma: a survival analysis of 137 consecutive patients.

AIM: To identify prognostic factors for long-term outcome for patients with hepatocellular carcinoma (HCC) after percutaneous microwave or radiofrequency ablation. MATERIALS AND METHODS: In total, 137 consecutive patients with HCC underwent microwave or radiofrequency ablation with curative intent; 16 possible prognostic factors were evaluated for their association with overall survival (OS) and disease-free survival (DFS) using univariate and multivariate analysis. RESULTS: The median OS and DFS were 27.0 months and 8.2 months, respectively. OS rates for all patients at 1, 2, 3, 4 and 5 years were 73.9%, 52.1%, 42.8%, 26.2% and 20.1%, respectively. DFS rates at 1, 2, 3 and 4 years were 38.1%, 21.9%, 18.8%, and 14.1%, respectively. Pretreatment serum alpha-fetoprotein (AFP) >200 ng/ml, pretreatment serum albumin 14 s were identified to be significant predictors for DFS by univariate analysis, and the former was the only independent predictor for DFS by multivariate analysis. CONCLUSION: Prognosis for patients with HCC after thermal ablation with curative intent was determined by treatment response to ablation, pretreatment serum AFP, and liver function reserve. Tumour response to treatment was the most predictive factor for long-term survival and was related to tumour size, thus careful selection of patients for ablation therapy is recommended.

Percutaneous thermal ablation for recurrent hepatocellular carcinoma after hepatectomy.

BACKGROUND: Tumour ablation using a thermal energy source has shown promising results, and is particularly suitable for recurrent hepatocellular carcinoma (HCC). The present study evaluated long-term outcomes after percutaneous thermal ablation for recurrent HCC following liver resection. METHODS: Radiofrequency ablation or microwave ablation was used to treat a total of 124 tumour nodules (0.9-7.0 cm in diameter) in 72 patients with recurrent HCC. RESULTS: Complete ablation of 119 (96.0 per cent) of 124 tumour nodules was achieved. There was no treatment-related death and the major complication rate was 4 per cent. During a mean(s.d.) follow-up of 27.9(17.8) months, local recurrence developed in 16 (13.6 per cent) of 118 successfully treated tumour nodules. Distant recurrence developed in 60 (85 per cent) of 71 patients, of whom 26 had repeat metachronous distant recurrence. With repeated ablation for both local and distant recurrence, the 1-, 3- and 5-year overall survival rates after initial ablation were 75, 43 and 18 per cent respectively. Patients with a serum alpha-fetoprotein level greater than 200 ng/ml before treatment had significantly poorer survival than those with a lower level (P = 0.034) and multivariate analysis identified preablation AFP level as an independent prognostic factor (P = 0.054). CONCLUSION: With their advantages of preservation of non-tumorous liver tissue and easy repetition, percutaneous thermal ablative therapies were particularly suitable for recurrent HCC and improved long-term survival.

Microwave tumour coagulation plus in situ treatment with cytokine-microparticles: induction of potent anti-residual tumour immunity.

After local microwave coagulation and subsequent intra-tumoural injection of microparticles encapsulating interleukin-2 and granulocyte-macrophage colony-stimulating factor, the anti-tumour efficacy against subcutaneous Lewis lung carcinoma in syngeneic mice was evaluated. This treatment elicited a potent systemic anti-tumour immunity that protected treated mice from re-challenge with the same tumour cells and caused the distal tumours in a bilateral tumour model to be rejected. Cytotoxicity assay indicated that both T- and natural killer cells acted as the effector cells in the anti-tumour immunity. These data highlight the feasibility of microwave-pre-treated in situ cancer vaccination for clinical use.

A tumour vaccine of fixed tumour fragments in a controlled-release vehicle with cytokines for therapy of hepatoma in mice.

BACKGROUND: Cytokines can be strong potentiators for a tumour vaccine, but they have very short life in vivo when administered as a solution. AIMS: To evaluate the slow release of interleukin 2 from a cytokine-vehicle in vitro and in vivo and to evaluate the anti-tumour activity of a new tumour vaccine in vivo. METHODS: The tumour vaccine was composed of formalin-fixed Hepa 1-6 hepatoma tissue fragments, tuberculin and a lipid based vehicle containing granulocyte-macrophage colony-stimulating factor and interleukin 2. The quantity of interleukin 2 release from the cytokine-vehicle in vitro and in vivo was determined by a proliferation assay with CTLL-2 cell line. Hepa 1-6 hepatoma model system with C57BL/6J mice was used to examine protective and therapeutic anti-tumour effect of the vaccine. RESULTS: Release of interleukin 2 from the cytokine-vehicle lasted 5 days in vitro and 3 days in vivo. The vaccine protected 67% of mice from a Hepa 1-6 cell challenge and had a therapeutic effect by prolonging the life span of mice bearing established Hepa 1-6 tumours of 5 mm in diameter. Of the treated mice, 20% became completely tumour-free. CONCLUSIONS: Formalin-fixed tumour fragments and cytokines in controlled-release vehicle are useful in the rational design of tumour vaccines.

[Relationship between hemolymph phenol oxidase and melanization of oocysts of Plasmodium yoelii in Anopheles stephensi].

OBJECTIVE: To explore the relationship between the hemolymph phenol oxidase and melanization of oocysts. METHODS: Anopheles stephensi-Plasmodium yoelii system was used to determine the activity of monophenol oxidase (MPO) and o-diphenol oxidase (o-DPO) in the hemolymph collected from 4 groups of mosquitoes by polyacrylamide gel electrophoresis (PAGE) followed by density scanning. The 4 groups of mosquitoes were: non-blood-fed (N), normal-blood-fed (B), infected-blood-fed (I) and nitroquine-administrated (D), respectively. RESULTS: No significant difference was found in the activities of MPO and o-DPO between groups N and B. The activities of MPO and o-DPO were not obviously modified in group I, but were significantly increased on day 10 and decreased on day 15 after blood feeding in the group D as compared with those in the groups N and B. CONCLUSION: The alteration in the mosquito hemolymph PO activity coincided at each time point with the melanization of Plasmodium yoelii oocysts.