Cancer multidrug resistance (MDR) impairs the therapeutic efficacy of various chemotherapeutics. Novel approaches, particularly the development of MDR reversal agents, are critically needed to address this challenge. This study demonstrates that tenacissoside I (TI), a compound isolated from Marsdenia tenacissima (Roxb.) Wight et Arn, traditionally used in clinical practice as an ethnic medicine for cancer treatment, exhibits significant MDR reversal effects in ABCB1-mediated MDR cancer cells. TI reversed the resistance of SW620/AD300 and KBV200 cells to doxorubicin (DOX) and paclitaxel (PAC) by downregulating ABCB1 expression and reducing ABCB1 drug transport function. Mechanistically, protein arginine methyltransferase 1 (PRMT1), whose expression correlates with poor prognosis and shows positive association with both ABCB1 and EGFR expressions in tumor tissues, was differentially expressed in TI-treated SW620/AD300 cells. SW620/AD300 and KBV200 cells exhibited elevated levels of EGFR asymmetric dimethylarginine (aDMA) and enhanced PRMT1-EGFR interaction compared to their parental cells. Moreover, TI-induced PRMT1 downregulation impaired PRMT1-mediated aDMA of EGFR, PRMT1-EGFR interaction, and EGFR downstream signaling in SW620/AD300 and KBV200 cells. These effects were significantly reversed by PRMT1 overexpression. Additionally, TI demonstrated resistance reversal to PAC in xenograft models without detectable toxicities. This study establishes TI's MDR reversal effect in ABCB1-mediated MDR human cancer cells through inhibition of PRMT1-mediated aDMA of EGFR, suggesting TI's potential as an MDR modulator for improving chemotherapy outcomes.
Humans ; Protein-Arginine N-Methyltransferases/antagonists & inhibitors* ; Drug Resistance, Neoplasm/drug effects* ; ErbB Receptors/genetics* ; Animals ; Cell Line, Tumor ; Drug Resistance, Multiple/drug effects* ; Methylation/drug effects* ; Saponins/administration & dosage* ; Mice ; Mice, Nude ; Mice, Inbred BALB C ; ATP Binding Cassette Transporter, Subfamily B/genetics* ; Doxorubicin/pharmacology* ; Paclitaxel/pharmacology* ; Female ; Repressor Proteins

Objective:To prepare a 177Lu labeled human epidermal growth factor receptor 2 (HER2) affibody 177Lu-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA)-maleimide (Mal)-cysteine (Cys)-ZHER 2: 342 ( 177Lu-NOTA-MZHER2 for short), and investigate its labeling process and anti-tumor properties. Methods:Two kinds of buffer systems (sodium acetate buffer system and sodium ascorbate buffer system) were investigated. The effects of pH value, precursor mass and reaction temperature on 177Lu labeling NOTA-MZHER2 were compared to obtain optimal labeling conditions. The radiochemical purity of labeled product was determined by instant thin-layer chromatography (ITLC), and its stabilities in PBS and plasma were observed. Human ovarian cancer cell line SKOV-3 was selected for cell internalization and cytotoxicity test to evaluate cell uptake and killing effect of 177Lu-NOTA-MZHER2. SKOV-3 tumor-bearing mice( n=3) were injected with 177Lu-NOTA-MZHER2, and microSPECT/CT imaging was performed. Another 40 tumor-bearing mice were divided into 22.2 MBq group (tail vein injection with probe of 22.2 MBq), control group (tail vein injection with PBS), low-dose group (tumor injection with probe of 3.7 MBq) and high-dose group (tumor injection with probe of 7.4 MBq). Tumor volume and mass of tumor-bearing mice were monitored after injection, and the anti-tumor effect and toxicity of probe were evaluated. Repeated measurement analysis of variance (Bonferroni method) was used to analyze the data. Results:The optimal labeling condition was 70-80 ℃ for 30 min in the system of sodium acetate buffer solution with pH=4 and precursor mass of 50 μg. Under these conditions, the labeling rate of 177Lu-NOTA-MZHER2 was (99.3±0.4)% and radiochemical purity was >99%. After 12 d in PBS and plasma, the radiochemical purities were (95.0±1.5)% and (95.0±2.1)%. Results of cell experiment showed that the internalization of 177Lu-NOTA-MZHER2 accounted for (29.02±3.50)% of the total uptake, and the survival rate of SKOV-3 cells was (48±6)% with the probe concerntration of 6×10 -3 Bq/L. SPECT imaging showed that 177Lu-NOTA-MZHER2 was still concentrated at the tumor site 96 h after injection with a dose of 18.5 MBq. Relative tumor volume (RTV) of tumor-bearing mice in 22.2 MBq group, high-dose group and low-dose group was significantly different from that in control group ( F=21.75, P<0.001). Twenty days after injection, RTV and relative body mass of the tumor-bearing mice in high-dose group were (140±7)% and (80±9)%, respectively. Compared with control group, high-dose group had obvious anti-tumor effect (both P<0.001). Conclusion:177Lu-NOTA-MZHER2 is successfully prepared, which is simple and efficient, and the probe has good anti-tumor effect.

Objective:To radiolabel hyperbranched polymer (HG)-modified liquid metal nanodroplet (LMND)@HG with 177Lu, and explore the radiotherapy sensitization effect on anti-breast cancer therapy. Methods:The ultrasonication method was used to prepare LMND@HG, and then 177LuCl 3 was mixed with LMND@HG to label 177Lu by alloying reactions. The labeling rate, plasma stability and cytotoxicity of 177Lu-LMND@HG were detected. Xenograft mouse model of breast cancer was constructed, and the tumor inhibition test was performed by an intratumoral injection. The tumor progression was monitored by in vivo imaging system. The mechanism of tumor inhibition was verified by immunohistochemistry and immunofluorescence assays. One-way analysis of variance, repeated measures analysis of variance, and the least significant difference t test were used to analyze the data. Results:177Lu was successfully labeled to LMND@HG with a high labeling efficiency >95%. The product did not require further purification and the plasma radiochemical purity was still higher than 95% after 5 d. The cytotoxicity test showed that a dose of 888 kBq (40 mg/L) 177Lu-LMND@HG had obvious toxicity to 4T1 cells, which was significantly lower than 177LuCl 3 (cell viabilities: (16.48±7.81)% vs (85.77±8.87)%; F=77.81, t=11.73, P<0.001) and LMND@HG ((46.53±5.75)%; t=6.20, P<0.001). The biological distribution results showed that 177Lu-LMND@HG was mainly distributed in tumor tissue 5 d after intratumoral injection. The results of the tumor inhibition experiment showed that 1.48 MBq 177Lu-LMND@HG could significantly inhibit the tumor growth compared with the 177LuCl 3 (tumor volume: (222.66±97.70) vs (789.13±245.04) mm 3;F=18.55, t=4.29, P=0.005). In vivo optical imaging of small animals showed that 1.48 MBq and 3.70 MBq 177Lu-LMND@HG both significantly inhibited the tumor growth. Immunofluorescence and immunohistochemical results showed that 177Lu-LMND@HG caused double-stranded DNA break, and suppressed the tumor growth by inhibiting cell proliferation and angiogenesis. Conclusions:A novel 177Lu-liquid metal-based reactive oxygen species (ROS) radiation sensitizer is successfully prepared in this study. The preparation method is efficient and convenient, and the product has high stability. 177Lu-LMND@HG shows an obvious radiotherapy sensitization effect on breast tumor-bearing mice.

OBJECTIVE To investigate the protective effect and potential mechanism of Arisaema Cum Bile on acute liver injury induced by carbon tetrachloride （CCl4） in mice. METHODS Fifty mice were randomly divided into normal group， model group， positive control group （Biphenyl diester dropping pills， 150 mg/kg）， Arisaema Cum Bile low-dose and high-dose groups （0.78， 2.34 g/kg）， with 10 mice in each group. The mice in each group were given relevant medicine intragastrically， once a day， for 7 consecutive days. Two hours after the last administration， those groups were given intraperitoneal injection of 0.2% CCl4-olive oil solution to induce acute liver injury model except for normal group. Seventeen hours after intraperitoneal injection， the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， interleukin-6 （IL-6）， tumor necrosis factor-α（TNF- α）， and malondialdehyde （MDA）， superoxide dismutase （SOD） in liver tissue were measured with kit method. The hepatic index was detected. The pathological changes of liver tissue were observed by HE staining， and the degree of liver injury was scored quantitatively. The mRNA expressions of TNF-α and IL-6 in liver tissue were detected by real-time fluorescence quantitative PCR； the protein expressions of Janus kinase 2 （JAK2）， signal transducer and activator of transcription protein 3 （STAT3） and nuclear factor-κB p65 （NF-κB p65） in liver tissue were detected by Western blot assay. RESULTS Compared with normal group，the levels of ALT， AST， IL-6， TNF-α and MDA， the hepatic index were increased significantly （P＜0.05）， while the level of SOD was decreased significantly （P＜0.05）； the mRNA E-mail：qhwang668@sina.com expressions of IL-6 and TNF-α， and the protein expressions of JAK2， STAT3 and NF-κB p65 were up-regulated significantly （P＜0.05）； the pathological observation of liver tissue showed that the structure of hepatic cord was seriously disordered， there were many inflammatory cells infiltration of liver cells， and the liver injury score was significantly increased （P＜0.05）. Compared with model group， pathological changes and above indexes in mice were improved significantly in Arisaema Cum Bile low-dose and high-dose groups （P＜0.05）. CONCLUSIONS Arisaema Cum Bile has a protective effect on CCl4-induced acute liver injury in mice， which may be related to the inhibition of inflammatory response mediated by JAK2/STAT3/NF-κB signal pathway and antioxidant stress.

Objective To evaluate the change of clinicopathological features and prognosis of papillary thyroid cancer over a 15-year period.Methods The clinicopathological features and outcomes of papillary thyroid cancer patients were analyzed in three groups according to the time of diagnosis:group Ⅰ (1997-2001),group Ⅱ (2002-2006),and group Ⅲ (2007-2011).Results As time advanced,the average age of papillary thyroid cancer patients increased,tumor stage,like size,extrathyroid invasion and lymph node metastasis decreased dramatically (P ＜ 0.01).The percentage of multifocality and bilaterality increased.The long-term follow up data (median follow up time was 6.6 years),indicated that the 15-year over all survival was 97.8％ and the 15-year disease-free survival was 90.2％.Tumor ≥3 cm,bilaterality,extrathyroid invasion,lymph node metastasis and AJCC stage were correlated with tumor recurrence.By multivariate COX-regression analysis only lymph node metastasis and bilaterality were independent risk factors.Conclusion The clinicopathological features of papillary thyroid cancer changed over 15 years,with the percentage of early-staged patients increased.Lymph node metastasis and bilaterality are two risk factors for tumor recurrence.

Melioidosis is a endemic infectious disease caused by Burkholderia pseudomallei, and is considered one of the major causes of fatal pneumonia and sepsis.This paper reports diagnosis and treatment course of one case pulmonary melioidosis, and reviews the related literatures, so to improve clinical workers'' understanding towards melioidosis, avoid misdiagnosis and missed diagnosis.

[ ABSTRACT] AIM:To compare the differences of the genome-wide methylation levels and methylated regions be-tween nasopharyngeal carcinoma ( NPC) cells in the same genetic background but different radiation resistance ( CNE-2 cells and CNE-2R cells).METHODS:Using the method which was developed by Doctor Zhao Cun-you, based on using methyl-sensitive restriction enzyme to measure the genome-wide methylation levels.In addition, MeDIP-Seq was used to analyze the methylated regions in 6 gene functional elements, including the upstream 2k sequence, 5’ UTR, coding se-quence, intron, 3’UTR and downstream 2k sequence, between CNE-2 cells and CNE-2R cells.RESULTS:The genome-wide methylation level was approximately 30%lower in CNE-2R cells than that in CNE-2 cells.No obvious difference on the amount of genes and the coverage of the peak in the 6 gene functional elements was observed.However, the methylation pattern of plentiful genes had altered in the gene function elements.CONCLUSION:The genome-wide methylation levels and methylated regions between NPC cells in the same genetic background but different radiation resistance were quite dif-ferent, indicating that the DNA methylation may be associated with NPC radioresistance.

Objective In order to prevent the infection of Acinetobacter baumannii and use antibiotics rationally,the clinical infection and drug resistant data of multi-drug resistance Acinetobacter baumannii (MRAB)detected in intensive care unit (ICU)of Beijing Friendship Hospital from 2011 to 2013were analyzed.Methods This study is a retrospective study.One hundred and eighty five strains of MRAB were collected from the patients in ICU from January 2011 to December 2013.Identificationand antibiotic susceptibility of strains were determined with Vitek-2 Compact automatic bacteria identification system.The annual infection rate of MRAB was counted.PCR was used to detect the resistance genes.The clinical features of the patients with MRAB were analyzed.The average age,acute physiology and chronic health evaluation (APACHE) Ⅱ score,duration in ICU and mortality ratio of the MRAB patients were compared with the patients without MRAB.Rank-sum test was used to analyze the average age,APACHE Ⅱ score and duration in ICU.Chi-squared test was used to analyze the mortality ratio and annual infection rate.Results The average age [(67 ± 17)vs (59-± 19) years old,Z =-5.365,P =0],APACHE Ⅱ score [(25.68±7.93) vs (17.62±8.39),Z=-14.821,P=0],duration in ICU [(27 ±29) vs (5 ±8) d,Z =-4.342,P =0] and mortality ratio [10.82％ (53/185) vs 28.65％ (147/1 359),x2 =45.92,P =0] of the patients infected by MRAB were significantly higher than those without the infection.The MRAB was found mostly in sputum and bronchial precipitates (83.78％,155/185).Though detection rate reduced yearly and there was a significant reduction in 2013 compared with 2011 [11.07％ (69/469) vs 8.37％ (52/621),x2 =8.755,P =0.003],the drug resistant rate was in high level and did not show any change in the 3 years.OXA-23 and OXA-51 were detected in all MRAB.Conclusions The main drug resistant mechanism of MRAB in ICU is related to OXA-23.More active methods of coutrol and prevention of MRAB should be used in elderly aud severely pneumonic patients.Intensive disinfection and isolation measures can decrease MRAB detection rate.Combined antibiotics should be used in patients with MRAB infection.

Objective To evaluate extracellular and intracellular IFNγ and IL-4 levels in assessing Th1/Th2 balance in children with chronic hepatitis B (CHB) and asymptomatic carriers.Methods Fiftyfour hospitalized children including 23 CHB patients and 31 asymptomatic carriers were collected from May 2007 to February 2009.Thirty-four healthy children were served as control.Serum IFNγ and IL-4 levels were measured by enzyme-linked immunosorbent assay (ELISA) and intracellular IFNγand IL-4 levels were detected by flow cytometer.Analysis of variance (for homogenous variance) and Kruskal-Wallis test (for non-homogenous variance) were performed.Results The differences on extracellular IFNγ, IL-4 and Th1/Th2 among CHB, asymptomatic carriers and control groups were not statistically significant (F=0.342, 0.020 and 0.507, P ＞ 0.05); while the intracellular IFNγlevels were (7.68 ± 4.62), (11.71 ±4.36) and (13.61 ±6.71) μg/mL, and Th1/Th2 ratios were 0.96 ±0.30, 1.67 ±0.76 and 2.11 ± 1.12in three groups respectively (F=0.255 and 0.140, P ＜ 0.05 or ＜ 0.01).The differences in intracellular IL-4 levels among three groups were not significant (F=0.425, P ＞ 0.05).Conclusions Cytokine balance is affected in CHB children and asymptomatic carriers, and flow cytometry analysis is considered as a better method in evaluating the status of Th1/Th2 balance.

Objective To evaluate the safety and efficacy of mesohepatectomy for treating centrally located liver tumors. Methods The clinical data of 9 cases of centrally located liver tumor treated in our hospital with mesohepatectomy were retrospectively analyzed. Meanwhile, previous reports in Chinese and English on mesohepatectomy for treating centrally located liver tumors were reviewed.Results In the current series, perioperative mortality and morbidity rates were 0 and 66.6%, respectively. Eight patients were alive during a follow-up of 3-38 months. Twenty clinical trials were included in our systematic review. Four were retrospective non-randomized trials comparing central hepatectomy with lobar or extended hepatectomy. The surgical mortality rate of mesohepatectomy was 0% ～7. 4 %. Frequent complications were bile leakage (0. 4% ～ 18. 5 % ), pleural effusion (5. 7 % ～ 23.5 % ), ascites ( 1.9 % ～ 11.6 % ) and pneumonia ( 1.7 % ～ 12.5 % ). No differences in perioperative morbidity and early complication rate were found between the mesohepatectomy group and lobar or extended hepatectomy group in all four non-randomized studies. Two studies revealed that the overall survival rate and disease-free survival of patients with hepatocellular carcinoma were similar between the 2 groups. Conclusion Mesohepatectomy is a safe and effective operative procedure for the treatment of centrally located liver tumors.