Toripalimab plus axitinib versus sunitinib as first-line treatment for advanced renal cell carcinoma: RENOTORCH, a randomized, open-label, phase III study.

BACKGROUND: Immune checkpoint inhibitors in combination with tyrosine kinase inhibitors are standard treatments for advanced clear cell renal cell carcinoma (RCC). This phase III RENOTORCH study compared the efficacy and safety of toripalimab plus axitinib versus sunitinib for the first-line treatment of patients with intermediate-/poor-risk advanced RCC. PATIENTS AND METHODS: Patients with intermediate-/poor-risk unresectable or metastatic RCC were randomized in a ratio of 1 : 1 to receive toripalimab (240 mg intravenously once every 3 weeks) plus axitinib (5 mg orally twice daily) or sunitinib [50 mg orally once daily for 4 weeks (6-week cycle) or 2 weeks (3-week cycle)]. The primary endpoint was progression-free survival (PFS) assessed by an independent review committee (IRC). The secondary endpoints were investigator-assessed PFS, overall response rate (ORR), overall survival (OS), and safety. RESULTS: A total of 421 patients were randomized to receive toripalimab plus axitinib (n = 210) or sunitinib (n = 211). With a median follow-up of 14.6 months, toripalimab plus axitinib significantly reduced the risk of disease progression or death by 35% compared with sunitinib as assessed by an IRC [hazard ratio (HR) 0.65, 95% confidence interval (CI) 0.49-0.86; P = 0.0028]. The median PFS was 18.0 months in the toripalimab-axitinib group, whereas it was 9.8 months in the sunitinib group. The IRC-assessed ORR was significantly higher in the toripalimab-axitinib group compared with the sunitinib group (56.7% versus 30.8%; P < 0.0001). An OS trend favoring toripalimab plus axitinib was also observed (HR 0.61, 95% CI 0.40-0.92). Treatment-related grade ≥3 adverse events occurred in 61.5% of patients in the toripalimab-axitinib group and 58.6% of patients in the sunitinib group. CONCLUSION: In patients with previously untreated intermediate-/poor-risk advanced RCC, toripalimab plus axitinib provided significantly longer PFS and higher ORR than sunitinib and had a manageable safety profile TRIAL REGISTRATION: ClinicalTrials.gov NCT04394975.

[Comparison of diagnostic value between renal cyst index and Bosniak classification in cystic renal masses].

A total of 106 patients who were diagnosed with Bosniak catergory ⅡF or Ⅲ cystic renal masses (CRM) and underwent surgery in Zhongshan-Xuhui Hospital and Xuhui District Central Hospital between January 2018 and December 2019 were retrospectively analyzed. The diagnostic accuracy of renal cyst index (RCI) and Bosniak classification system was compared by analyzing the relevant parameters including the sensitivity and specificity. There were 62 males and 44 females, with a median age of 56 years old. Among the 106 CRM, 72 were benign and 34 were malignant. The sensitivity and specificity of RCI was 94.12% and 81.94%, respectively, while the sensitivity and specificity of Bosniak classification system was 73.53% and 80.56%, respectively. Chi-square test revealed that the sensitivity of RCI was significantly higher than that of Bosniak classification system (P=0.023). The current study indicates that RCI is a simple and feasible method which can provide quantitative evaluation for predicting characteristics of CRM.

[Role and potential value of circular RNAs in the occurrence of primary hepatic cancer].

Circular RNA is a class of non-coding RNAs, which are covalently closed and circular at both ends, showing dissimilar characteristics from linear RNA. Several studies have shown that circular RNAs play an important role in the occurrence and development of primary hepatic cancer. By combining with the latest research progress of this field at home and abroad, we summarized the mechanism regulating the occurrence and development of liver cancer, abnormal expression, and as potential molecular markers for disease diagnosis and treatment.

[Application of mini-flank open nephron sparing surgery via retroperitoneal route for centrally located renal tumor treatment: a single center experience].

Objective: To investigate the safety and feasibility of mini-flank open nephron sparing surgery (MI-OPN) via retroperitoneal route for the treatment of centrally located renal tumor. Methods: From May 2013 to April 2015, twenty-four cases of centrally located renal tumor were treated with MI-OPN via retroperitoneal route in Zhongshan Hospital. All cases were included in this study with whose clinical data and long term follow-up information retrospectively analyzed. Results: With the assistance of intraoperative ultrasonography to confirm tumor location and boundary, MI-OPN was successfully performed in all cases. Mean tumor maximum diameter was 3.3±0.6 cm, mean operation time was 113±16 minutes, mean ischemia time was 31±6 min, and mean estimated blood loss was 102±46 ml. Mean postoperative hospital stay was 5.0±0.8 days, postoperative complication was found in one patient (4%). The mean pre- and postoperative serum creatinine were 77.1±20.1 µmol/L and 90.3±20.0 µmol/L. Pathological examination confirmed negative surgical margin in all cases, with 18 cases of clear cell renal cell carcinoma, 2 cases of papillary renal cell carcinoma, 2 cases of chromophobe renal carcinoma, 1 case of renal oncocytoma and 1 case of renal angiomyolipoma. In up to 12-36 months postoperative follow-up, no local recurrence or systemic progression was witnessed. Conclusions: For the treatment of centrally localized renal tumor, MI-OPN via retroperitoneal route is a safe and feasible operation method. Importantly, rupture of the tumor capsule was effectively avoided during tumor resection with the assistance of ultrasonic position-setting. Furthermore, incidence of severe postoperative complications such as bleeding and damage of collection system were not found since surgical wound of kidney sewn tightly and finely. The last but not the least, by placing ice slush in retroperitoneal cavity, impairment of renal function caused by renal artery clamping can be alleviated due to decreased metabolism.

[Outcomes of mini-flank incision for open partial nephrectomy for stage T1b renal tumor].

Objective: To evaluate the safety and efficacy of mini-flank incision for open partial nephrectomy for stage T1b renal tumor. Methods: The data of patients with stage T1b renal tumor who underwent mini-flank incision for open partial nephrectomy between January 2010 to September 2015 were retrospectively reviewed. The Nephron-sparing surgery (NSS) was performed through mini-flank supra-12th rib incision under general anesthesia. Results: A total of 47 patients(31 male and 16 female) were enrolled in our study. The median age was 40 years (range 22-67 years). The Zhongshan Score(ZS score) of renal tumors was 6 in 5 cases, 7 in 13 cases, 8 in 12 cases, 9 in 5 cases, 10 in 6 cases, 11 in 2 cases, 12 in 2 cases, 13 in 2 cases. The length of incision was from 7 cm to 9 cm, with an average of 8.1 cm. The operative time was from 70 min to 150 min, with an average of 96 min. The blood loss was from 50 ml to 600 ml, with an average of 135 ml. The warm ischemia time was from 20 min to 35 min, with an average of 28 min. All of the surgery margin were negative. One patient had fluid in surgical region and relieved after the drainage, and one patient had acute myocardial infarction. The hospital stay time was from 5 d to 14 d, with an average of 8 d. The pathological diagnosis included clear cell carcinoma in 37 cases, multilocular cystic renal carcinoma in 1 case, chromophobe cell tumor in 4 cases, and papillary carcinoma in 5 cases. The mean preoperative serum creatinine level was 87 µmol/L(48-150 µmol/L) and with a mean of 91 µmol/L(52-148 µmol/L) at 3 month follow-up after surgery, and there was no difference between the preoperative and postoperative period(P>0.05). A total of 45 out of 47 patients were followed up for 36 to 78 months, with an average of 60 months, and no one had recurrence or metastasis during follow-up. Conclusion: Mini-flank incision for open partial nephrectomy for renal tumor with stage T1b is safe and effective, which is worthy of promotion and application for small incision and quick recovery.

Differential diagnosis between acinic cell carcinoma and pleomorphic adenoma using the quantitative parameters of contrast-enhanced ultrasound.

OBJECTIVE: To investigate the value of differential diagnosis between acinic cell carcinoma (ACC) and pleomorphic adenoma (PA) using the quantitative parameters of contrast-enhanced ultrasound (CEUS). PATIENTS AND METHODS: Twenty-two ACC and 98 PA were retrospectively analyzed. These patients had been examined via routine pre-surgical two-dimensional ultrasound and CEUS. The examination results were confirmed by biopsy pathology. Qontrast 4.0 imaging analysis software was applied to obtain the maximum intensity (PEAK), time to peak (TTP), regional blood volume (RBV), regional blood flow (RBF), maximum signal intensity (SImax) and mean signal intensity (SImean) through quantitative analysis. The differences between ACC and PA were compared regarding the conventional ultrasound images and the quantitative parameters of CEUS. ROC curves were drawn to evaluate the diagnostic value of these parameters. RESULTS: There were no statistically significant differences between salivary gland ACC and PA in the manifestations of conventional two-dimensional ultrasound examination regarding morphology, internal echo and the boundary (p > 0.05). However, there were significant differences in PEAK, RBV, RBF, SImax and SImean between ACC and PA (p < 0.05). Additionally, the five quantitative parameters of CEUS were all highly accurate diagnostic indicators. The maximum area under the curve of each parameter was 0.888, sensitivity 72.6%, specificity 90.9% and accuracy 81.8%. CONCLUSIONS: The quantitative parameters of CEUS are helpful for differentially diagnosing salivary ACC and PA.

[Expression of CD45 in newly diagnosed multiple myeloma and the relationship with prognosis].

Objective: To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis. Methods: This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45(+)/CD45(-) groups. Results: ①The CD45(+) group was 33 cases (25.38%) , and CD45(-) group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45(+) and CD45(-)group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45(+) and CD45(-) group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45(+) group and CD45(-) group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ(2)=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ(2)=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 (+) group and CD45(-) group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ(2)=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ(2)=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 µmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors. Conclusion: CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45(+) MM patients.

Chitosan-Based Extrafibrillar Demineralization for Dentin Bonding.

Instability of resin-dentin bonds is the Achilles' heel of adhesive dentistry. To address this problem, a chelate-and-rinse extrafibrillar dentin demineralization strategy has been developed that keeps intrafibrillar minerals within collagen fibrils intact to prevent activation of endogenous proteases that are responsible for collagen degradation within hybrid layers. The objective of the present study was to evaluate the potential of using chitosan >40 kDa as an antimicrobial extrafibrillar dentin-chelating agent to enhance bond durability. Transmission electron microscopy provided evidence for retention of intrafibrillar minerals and smear plugs in dentin conditioned with 1 wt% chitosan. Analyzed by Kruskal-Wallis analysis of variance, Dunn's statistic, and separate Mann-Whitney tests, tensile bond strengths to wet- and dry-bonded dentin indicated that chelating dentin with chitosan for 60 s prior to bonding did not result in a significant decline in resin-dentin bond strength when compared with that of phosphoric acid etching ( P > 0.05). Gelatinolytic activity within the hybrid layers was examined via in situ zymography after 24-h storage or after thermomechanical cycling and analyzed with 3-factor analysis of variance. After 24 h, enzymatic activity was detected only within completely demineralized phosphoric acid-etched dentin, with values derived from dry bonding significantly higher than those derived from wet bonding ( P < 0.05). Negligible fluorescence was detected within hybrid layers when dentin was conditioned with chitosan, even after thermomechanical cycling, as compared with the controls. Reduction in water permeability in chitosan-conditioned dentin, attributed to smear plug retention, also fostered long-term bond stability. Antibacterial testing performed with live/dead staining indicated that the acetic acid-solubilized chitosan possessed antibacterial activities against 3 single-species biofilms: Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis. Taken together, the new chitosan-based extrafibrillar demineralization strategy retains intrafibrillar minerals, reduces endogenous protease-initiated collagen degradation, prevents water permeation within hybrid layers, and kills bacteria on dentin surfaces, which are crucial factors for enhancing resin-dentin bond durability.

[Analysis of clinical characteristics and prognostic factors in patients with non-Hodgkin lymphoma and HBV infection].

Objective: To explore the clinical characteristics and prognostic factors of the patients with non-Hodgkin's Lymphoma (NHL) complicated with HBV infection, so as to provide a basis for clinical accurate diagnosis and prognosis evaluation. Methods: The data of 313 newly diagnosed NHL patients from August 2012 to July 2016 were collected. The HBV serological markers were detected by ELISA, and HBV DNA was quantified by full automatic microparticle chemiluminescence immunoassay (≥1×10(5) copies/ml as high copy group, 1×10(3)-<1×10(5) copies/ml as low copy group). The relationship between HBV infection and prognosis was analyzed combined with the clinical features of the patients, and the HBV detection rate was compared with that of the common population (from the national HBV sero epidemiological data). Results: ①The positive rate of HBsAg in NHL patients was 12.5% (39/313), which was higher than 7.2% in the general population (χ(2)=14.596, P<0.001). HBV infection in the past (HBsAg negative but HBcAb positive) in 114 cases (36.4%), the incidence was slightly higher than that in the general population (34.1%). ②Compared HBsAg positive group with the negative group, the proportion of B cell type (87.2% vs 70.3%, P=0.027), Ann Arbor stage Ⅲ-Ⅳ(69.2% vs 34.6%, P<0.001), IPI score 3-5 (74.4% vs 50%, P=0.004), LDH level (79.5% vs 47.8%, P<0.001) and liver involvement (45.5% vs 31.7%, P=0.006) were all higher. The difference was statistically significant. ③Compared the HBV infected group (114 cases) with the non-infected group (160 cases), the difference had statistical significance in the proportion of Ann Arbor stage Ⅲ-Ⅳ (P=0.023) and IPI score 3-5 scores P=0.035). ④Compared HBV DNA positive group (30 cases) with negative group (71 cases), Ann Arbor stage Ⅲ-Ⅳ (P=0.011), IPI score 3-5 score (P=0.030), LDH level (P=0.025) and liver involvement (P<0.001) in the proportion of patients had statistical significance. The positive patients were divided into HBV DNA high and low copy groups with 1×10(5) copies of /ml as the boundary. The results showed that there was no statistical difference between the two groups (P>0.05). Conclusions: The HBV infection rate of NHL patients is significantly higher than that of the general population, and HBV infection is more closely related to B cell type NHL. Patients with HBV infection and HBV DNA positive had late Ann Arbor stage, high IPI score, high LDH level and liver involvement, and the prognosis is poor.

[Prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric acute myeloid leukemia].

Objective: To investigate the prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric patients with t (8;21) acute myeloid leukemia (AML) . Methods: The clinical features and RUNX1-RUNX1T1 transcript levels of 55 pediatric t (8;21) AML patients, newly diagnosed from Jan. 2010 to Apr. 2016, were analyzed retrospectively. The relationship between the minimal residual disease (MRD) and prognosis was analysed by dynamic monitoring of RUNX1-RUNX1T1 transcript levels using real-time quantitative PCR (RQ-PCR) technology. Results: The RUNX1-RUNX1T1 transcript levels in bone marrow cells at diagnosis was not related to relapse. After one course of induction therapy, patients with a more than 2 Log reduction of RUNX1-RUNX1T1 transcript levels (>2 Log) had lower 5 years cumulative incidence of relapse (CIR) [ (24.3±8.4) % vs (52.6±9.7) %, χ(2)=9.046, P=0.003], relapse-free survival (RFS) [ (71.6±12.7) % vs (48.1±13.2) %, χ(2)=5.814, P=0.016], and better overall survival (OS) [ (76.9±12.5) % vs (48.9±14.7) %, χ(2)=6.346, P=0.012], compared to patients with a less than 2 Log reduction (a<2 Log) . Multivariate Cox survival analysis suggested that a>2 Log reduction in RUNX1-RUNX1T1 transcript levels after a course of induction therapy was an independent prognostic factor for RFS (HR=0.263, 95%CI 0.081-0.851, P=0.026) and OS (HR=0.214, 95% CI 0.057-0.808, P=0.023) . During consolidation therapy and follow-up period, molecular relapse of 16 cases and hematologic relapse of 13 cases were identified by continuous dynamic monitoring of RUNX1-RUNX1T1 transcript levels, with a median interval of 4.0 (1.5-5.8) months from the molecular relapse to hematologic relapse. 2 cases of molecular relapse who received timely allogeneic hematopoietic stem cell transplantation did not experience hematologic relapse. Conclusion: Dynamic monitoring RUNX1-RUNX1T1 transcript levels by RQ-PCR technique can subdivide patients into relatively low and high risk group, early screen patients at high risk of relapse and provide a scientific basis for precision stratification and risk-adapted therapy for pediatric t (8;21) AML children.

[The analysis of prognosis-associated factors in adults with acute promyelocytic leukemia].

Objective: To explore the prognostic value of CD34, CD2, CD56 expressions and FLT3-ITD mutation in adults with acute promyelocytic leukemia (APL) . Methods: The immuno-phenotypic and molecular characteristics of 137 adult patients with APL (from January 2010 to March 2016, in Henan Provincial People's Hospital) were investigated. And the relationships between CD34, CD2, CD56 expressions, FLT3-ITD mutation and the outcomes of high WBC counts at onset, complete remission (CR) rate, early mortality, relapse rate (RR) , overall survival (OS) , disease free survival (DFS) were explored. Results: ①Among the 137 patients, the positive ratios of CD34, CD2, CD56 expressions and mutation rate of FLT3-ITD were 26.3%, 25.5%, 10.2% and 17.5%, respectively. The morbidities of positive CD34, CD2, CD56 expressions and FLT3-ITD mutation in the high-risk group were 43.2%, 47.7%, 18.2% and 27.3% respectively, while those in the low-/intermediate-risk groups were 18.3%, 15.1%, 6.5% and 12.9%, respectively (P<0.05) . ②At a median follow-up of 41 months, the total CR rate of the 137 adults APL patients was 96.9%, early mortality 6.6% and relapse rate 7.3% respectively. And RR of positive CD34 or CD2 expression patients was higher than negative CD34/CD2 expression ones (18.8% vs 3.3%, χ(2)=8.462, P=0.004; 16.1% vs 4.3%, χ(2)=4.382, P=0.028, respectively) . In addition, the early mortality of patients with positive CD56 expression or FLT3-ITD mutation was extremely higher than in negative ones (21.4% vs 4.9%, χ(2)=5.610, P=0.018; 16.7% vs 4.4%, χ(2)=4.833, P=0.028, respectively) . ③The whole OS and DFS were 88.3% and 84.7%, respectively. Wherein, OS and DFS in patients with CD34(+), CD56(+) or FLT3-ITD mutation were worse (P<0.05) . Conclusions: Positive CD34, CD2, CD56 expression and FLT3-ITD mutation were latent poor prognostic factors in adults with APL.

[Efficacy of combination of ATRA, ATO and anthracyclines induction therapy in patients with acute promyelocytic leukemia].

Objective: To explore the efficacies of regimens of three-drug induction therapy (ATRA+ATO+anthracyclines) versus two-drug induction therapy (ATRA+ATO) in patients with acute promyelocytic leukemia (APL). Methods: Of 184 patients diagnosed with APL from January 2009 to March 2016, 58 patients underwent three-drug induction therapy, while the rest were treated with two-drug induction therapy. Three-drug induction therapy was of ATRA (20 mg·m(-2)·d(-1), d(1-28)) + ATO (0.16 mg·kg(-1)·d(-1), d(1-28)) + Idarubicin (8 mg·m(-2)·d(-1), d(3-5)) /daunorubicin (40 mg·m(-2)·d(-1), d(3-5)) , while two-drug induction therapy ATRA+ATO with the same doses and methods as above. Of 184 cases, 69 cases accompanied with WBC counts>10×10(9)/L, 115 cases with WBC counts≤10×10(9)/L at onset. Results: ①Short-term efficacy: After one cycle induction therapy, the rates of hematologic remission, genetic remission, molecular remission and induced differentiation syndrome (DS) in three-drug regimen group were 98.3%, 87.9%, 72.4% and 0 respectively, while those in two-drug regimen group were 87.3%, 65.9%, 51.6% and 12.7% respectively. In patients with WBC >10×10(9)/L, DS rate and early mortality in three-drug regimen group were lower than in two-drug regimen group (0 vs 15.6%, 4.2% vs 15.6%, respectively). In patients with WBC≤10×10(9)/L, DS rate in three-drug regimen group was also lower than in two-drug regimen group (0 vs 12.3%) , but there were no statistical differences in terms of relapse and early mortality. ② Long-term efficacy: The relapse rate, overall survival (OS) and disease free survival (DFS) in three-drug regimen group were 0, 98.5%, 96.6% respectively, while those in two-drug regimen group were 8.6%, 86.5% and 84.1% respectively; the advantages of three-drug over two-drug regimen, especially in cases of WBC >10×10(9)/L were observed. ③ Side effects: the incidences of gastrointestinal reaction, liver dysfunction, myocardial damage and headache in three-drug regimen group hardly increased. Conclusion: The efficacies of three-drug induction therapy were superior to two-drug one.

The effects of all-trans-retinoic acid on cell cycle and alkaline phosphatase activity in pancreatic cancer cells.

Pancreatic cancer is one of the tumors with the highest mortality, poorly responding to available chemotherapeutic agents. The objective of this study was to study the anticancer effects of all-trans retinoid acid, a functional form of vitamin A, on pancreatic cancer cells. Human pancreatic cancer MiaPaCa-2 cells were treated with 1, 5, 10, 20, 30, 40 and 50 microM ATRA for 1, 2, 3, 4, 5 or 6 d, respectively. Cell growth was determined by MTT viability assay. The cell cycle distribution and the alkaline phosphatase (ALP) activity were analyzed by flow cytometry and chemical analyzer, respectively. The results show that ATRA significantly inhibited the growth of MiaPaCa-2 cells at 40 and 50 microM. ATRA arrested pancreatic cancer cells at G0/G1 phase. The sub-G1 peak and DNA fragmentation were observed. There were time and dose dependent increases in alkaline phosphatase activity (ALP), an indicator of cell differentiation, upon treatment with ATRA when compared to controls. In conclusion, ATRA has an inhibitory effect on the cell growth of MiaPaCa-2, and its tumor suppressive effect is by means of cell cycle arrest and apoptosis induction.

[Effects of exogenous enzymes on the degradation of adhesive-dentin interfaces].

OBJECTIVE: To compare the effects of exogenous enzymes on the degradation of adhesive-dentin interface. METHODS: Forty molars were sectioned to expose the middle-coronal dentin surface and randomly divided into two adhesive systems: an etch-and-rinse adhesive Adper Single Bond 2 and a self-etching adhesive G-Bond. After composite building up, the specimens were then randomly assigned to four groups(n=5 for each group)as follows: group 1, 24 h of water storage(the control group); group 2, six months of water storage; group 3, twelve weeks storage in artificial saliva containing clostridium histolyticum collagenase; group 4, twelve weeks storage in artificial saliva containing cholesterolesterase. The microtensile bond strengths(MTBS)were then tested. The failure modes and nanoleakage were analyzed. RESULTS: After aging treatments, the three aging groups showed significantly lower MTBS compared with the control group in both adhesive systems(P<0.05). For etch-and-rinse adhesive Adper Single Bond 2, the MTBS of group 3([19.6±3.5]MPa)was lower than that of group 2([23.4±4.2]MPa)and group 4([24.2±4.2]MPa)(P<0.05). For self-etching adhesive G-Bond, there was no difference on MTBS among different aging groups(P>0.05). SEM observation showed that, compared with the control group, water storage(group 2)and the exogenous enzymes(group 3 and 4)increased the nanoleakage expression(silver deposition)of both adhesive systems. Adhesive failure was the predominant fracture modes in all groups. CONCLUSIONS: Storage in artificial saliva containing clostridium histolyticum collagenase or cholesterol esterase could be used to accelerate the degradation process of adhesive-dentine interface.

Structural alterations of sugar chains in urine fibronectin from bladder cancer patients and its enzymatic mechanism.

PURPOSE: To study the structural alterations of asparagine-linked sugar chains (N-glycans) on urine fibronectin (Fn) from bladder cancer (BCa) patients and its enzymatic mechanism. METHODS: Eight pairs of urine samples from eight BCa patients pre-operation and 3 months post-operation (which proved to be normal) were collected, and the Fn in the urine samples was purified with an anti-Fn antibody affinity column. Different lectins labeled with horseradish peroxidase (HRP) were used as probes to bind the glycans of purified Fn immobilized on membrane. Enhanced chemiluminescence (ECL) reagent was adopted to estimate the activity of the bound HRP as a measure of the binding affinity of the Fn glycans to lectins, and expressed as luminescent light units (LLU). The enzymatic mechanism of the structural alteration of N-glycans in BCa Fn was studied by determination of GnT activities using the HPLC method and fluorescent-labeled substrate. RESULTS: The mean LLU of BCa Fn was only 18.1% of the normal samples when Con A-HRP were used as probes, while the mean LLU of the BCa group was 3.34 times and 3.26 times higher than normal for the DSA-HRP and WGA-HRP probes, respectively. The individual data of the patients did not overlap between the BCa sample and normal counterparts, indicating that the positive rates were 100%, regardless of which lectin-HRP was used. These results reveal that the antennary number and bisecting GlcNAc structure are increased in the N-glycans of urine Fn from BCa assessed according to the binding specificity of ConA, DSA, and WGA. In addition, the binding affinities of urine Fn with DSA and WGA were correlated to pathological stage, and the affinity of Fn with WGA was also correlated with pathological grade. The results of GnT determination showed that GnT-III, IV, and V in BCa tissues increased by 34.0, 18.1, and 1.6 times, respectively, in normal bladder tissues which were at least 5 cm away from the BCa of the same bladder. These findings were compatible with the structural changes of N-glycans in BCa Fn, since GnT-III and GnT-IV/V are responsible for the synthesis of bisecting GlcNAc and the increase of antennary number in N-glycans, respectively. CONCLUSIONS: (1) The highest elevation of GnT-III and the close relationship between the WGA binding of BCa Fn with the pathological stage and grade of BCa indicate that the increase of bisecting GlcNAc in N-linked glycans contributes more to the malignant behavior of BCa than the increase of GnT-IV, GnT-V, and the antennary number. (2) The correlation of altered activities of bladder GnTs with the abnormal structures of urine Fn in BCa patients indicates that the urine Fn is synthesized in the bladder. (3) The lectin-HRP assay for analyzing the structure of N-glycans in urine Fn may be used as a simple and accurate diagnosis method for BCa in the future.

Biphasic effect of daidzein on cell growth of human colon cancer cells.

Colorectal cancer is one of the most common cancers in the world, poorly responding to available chemotherapeutic agents. To investigate whether natural molecules can inhibit colon cancer progression, we investigated a principle phytoestrogen found in soybean known as daidzein, and determined its effects on the human colon cancer cell line LoVo. LoVo cells were treated with 0.1, 1, 5, 10, 50 and 100 microM daidzein for 2, 3, 4 or 5 d. The results indicated that daidzein stimulated the growth of LoVo cells at 0.1 and 1 microM whereas at higher concentrations (10, 50 and 100 microM) cell growth was inhibited in a dose-dependent manner. Treatment of daidzein at 10, 50 and 100 microM resulted in cell cycle arrest at G0/G1 phase, DNA fragmentation and increases in caspase-3 activity. There were no changes in alkaline phosphatase activity (ALP), an indicator of cell differentiation, upon treatment with daidzein when compared to controls. These results indicate that daidzein has a biphasic effect on LoVo cell growth and its tumor suppressive effect is by means of cell cycle arrest and apoptosis but not through cell differentiation.

[Analyses of chromosomal karyotypes and cytogenetic variations of animal cell lines].

After the master cell stock(mcs) and working cell bank of more than 30 different strains of 7 animal kidney cell lines (F-81 or CRFK cell line, MDCK cell line, Vero or Vero-2 cell line, MA-104 cell line and BHK-21 cell line) were established in China, the chromosomal number variations and structural aberrations of the above lines, primary feline or canine kidney cell (FKC or CKC) and HeLa cell line were investigated and their karyotypes of routine or Giemsa chromosomal bands were analyzed. The carcinogenesis or tumorigenicity testing of these cells in about 700 nude mice and for colony formation in soft agar (SA) and for agglutination under different concentration of plant lectins was carried out. Both tumorigenicity-negative strains of F-81, CRFK, Vero or Vero-2 lines and very-low-tumorigenicity strains of MDCK line were successfully selected and evaluated for the production of canine or feline combination viral vaccines, which are free of infectious agents, and described with respect to cytogenetic characteristics and tumorigenicity. Rate of modal chromosome number represents the ratio of cell number having modal chromosome number to all the split cell number analyzed at random. Rate of difference represents the ratio of difference of the rate of modal chromosome number between mcs (master cell stock) + n and mcs passages. The chromosomal analysis results showed that the ratio of difference of the rate of modal chromosome number between mcs + n and mcs passages was not more than 5%-15% and the structure aberrations was generally 0%-3%, not more than 5%-10%, thus the hereditary character of cell lines is comparatively stable without significant difference between different passages. The genetic characteristics of chromosomal number of cell lines determines their tumorigenicity, but it is species specific. Experimental models were established for the researches on the prevention and prophylaxis of malignant tumors or cancers and their genetically biological characteristics. Tests showed that there was correlation among cell line chromosome number variations, anchorage independence in soft agar, agglutination under plant lectins and tumor-forming ability in nude mice. Since testing in vitro is more economic, simpler, faster, and is thought to be reliable, we recommend plant lectins followed by SA or analysis of karyotypes as the initial means for monitoring tumorigenicity of animal cell line in nude mice.

(125)I particle implantation combined with chemoradiotherapy to treat advanced pancreatic cancer.

OBJECTIVE: To evaluate the therapy effects of (125)I implantation combined with chemoradiotherapy on pancreatic cancer patients. METHODS: 30 patients with Stage III or IV pancreatic cancer were equally divided into two groups (control and treatment group). The patients in the treatment group (nine males, six females) received chemotherapy in the first week and (125)I implantation in the third week, followed by combined chemoradiotherapy in the fifth week. The patients in the control group (10 males, 5 females) received the same treatment except (125)I implantation. The therapy in the control group and treatment group was repeated every 4 weeks. RESULTS: The median conformal radiotherapy dose in the treatment group (30.62 Gy) was significantly lower than that in the control group (47.86 Gy). The total radiation dose was 88.71 ± 27.39 Gy, and the surface activity was 0.6 mCi in the treatment group. After treatment, the average tumour size decreased both in the treatment group [9.17 cm(2), 95% confidence interval (CI): 5.60-12.74, p < 0.001] and in the control group (4.54 cm(2), 95% CI: 2.74-6.35, p < 0.001). The median survival time in the treatment group was 14 months (95% CI: 12.215-14.785) and in the control group was 12 months (95% CI: 10.884-13.116). There was no statistical significance in survival rates between the two groups (χ(2) = 0.908, p = 0.341). CONCLUSION: (125)I implanted into tumour combined with chemoradiotherapy has higher local control rate of advanced pancreatic cancer than chemoradiotherapy. ADVANCES IN KNOWLEDGE: We combined chemoradiotherapy with (125)I implantation to treat advanced pancreatic cancer and obtained a higher local control rate and better quality of life than when using chemoradiatherapy alone.

Effectiveness and security of CT-guided percutaneous implantation of (125)I seeds in pancreatic carcinoma.

OBJECTIVE: To assess the effectiveness and security of CT-guided percutaneous implantation of iodine-125 ((125)I)-labelled seeds in pancreatic carcinoma. METHODS: A total of 36 patients (25 males and 11 females) with an average age of 57 years (range, 39-84 years) were enrolled and categorized into Stage III (27 cases) and Stage IV (9 cases) of pancreatic cancer. There were 3 tumours in the pancreatic head and 33 tumours in the pancreatic body or tail. The average diameter of the tumours was 37.1 mm (range, 15-65 mm). The implantation of (125)I seeds was performed by using 18-G needles (length, 150-200 mm) through the anterior, lateral and posterior approaches. Then, (125)I seeds were loaded and released into the lesions. RESULTS: Implantations were performed via the anterior (23 patients), lateral (9 patients) and posterior (4 patients) approaches. During implantation, 3-14 punctures were performed for each patient, and a total of 164 punctures were recorded. Meanwhile, a total of 657 seeds were implanted with an average of 25.27 (range, 12-50) seeds per patient, and the success rate was 100%. The activity of each seed ranged from 0.55 to 0.65 mCi. A main adverse event occurred in one puncture and minor events in seven punctures. No significant relationship between the punctures or adverse events was identified. No serious complication was detected after the implantations during follow-up visits. CONCLUSION: This study suggested that CT-guided percutaneous implantation of (125)I seeds in a pancreatic carcinoma was relatively safe and effective for treating unresectable pancreatic cancer. ADVANCES IN KNOWLEDGE: The CT-guided percutaneous implantation of (125)I seeds in unresectable pancreatic cancer showed highly successful rates without serious complications.