0.05).The expression of survivin mRNA and BCRP mRNA was correlated with therapeutic effect of chemotherapy(P

Objective To investigate the effect of deltaNp63(ΔNp63) silencing on the proliferation of pancreatic cancer PANC1 cells.Methods ΔNp63 mRNA level in 23 pairs of pancreatic cancer and adjacent tissue specimen was detected by real-time PCR, andΔNp63 protein in human normal pancreatic ductal cell line HPDE6-C7 and pancreatic cancer cell line PANC1, CFPAC1 and BXPC3 was detected by Western blot. PANC1 cells were transfectedΔNp63 specific siRNA (ΔNp63-siRNA ) and scramble siRNA ( Con-siRNA ) using liposome, and untransfected cells served as control.ΔNp63 mRNA and protein was detected by real-time PCR and Western blot to validate the silencing ofΔNp63 expression.MTT assay and BrdU method were used to detect the proliferation and DNA synthesis of transfected PANC1 cells.Results TheΔNp63 mRNA expression in pancreatic cancer tissues and matched adjacent normal tissues was 0.99 ± 0.07 and 0.70 ±0.07, respectively.ΔNp63 mRNA expression in pancreatic cancer tissue was significantly up-regulated compared with that in the normal tissue (P=0.0034).TheΔNp63 protein expression in HPDE6-C7, PANC1, CFPAC-1 and BxPC3 cells was 0.97 ±0.09,3.06 ±0.16,2.57 ±0.11 and 2.45 ±0.08, respectively.TheΔNp63 protein level in pancreatic cancer cells were higher than that in HPDE6-C7 cells (P<0.001).ΔNp63 mRNA level in control, Con-siRNA and ΔNp63-siRNA group was 0.97 ±0.07,0.97 ±0.07 and 0.28 ±0.03, respectively, andΔNp63 protein expression level was 0.97 ±0.06,1.00 ±0.10 and 0.26 ±0.03.The expression ofΔNp63 mRNA and protein inΔNp63-siRNA group were significantly down-regulated comparing with those in Con-siRNA group (P<0.01).Significant inhibition on cell proliferation was observed in ΔNp63-siRNA group, which was statistically different from that in control and Con-siRNA group.The A490 value (DNA synthesis) of control, Con-siRNA andΔNp63-siRNA group was 0.55 ±0.04, 0.56 ±0.01 and 0.55 ±0.00 at 24 h after transfection, and 0.84 ±0.05,0.87 ±0.07 and 0.71 ±0.05 at 48 h after transfection.The DNA synthesis inΔNp63-siRNA group was significantly down-regulated compared with that in control and Con-siRNA group (P<0.05).Conclusions Knockdown ofΔNp63 could greatly inhibit the proliferation and DNA synthesis of pancreatic cancer PANC1 cells.

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.

Objective:To investigate the effects of Mor-platin, a novel mitochondrial platinum complex, on proliferation and migration of human hepatoma carcinoma HepG2 cells. Methods:Cell counting kit-8 (CCK-8) assay was used to analyze cell proliferation of Mor-platin and classic anticancer drugs, particularly cisplatin, in HepG2 cells. A laser confocal microscope was used to observe whether Mor-platin can target mitochondria. The morphological changes in cellular mitochondria after treatment with Mor-platin were ob-served on a transmission electron microscope. Cell apoptosis was measured by flow cytometry, and cell invasion was evaluated by three-dimensional tumor spheroid model. Results:Mor-platin can inhibit cell proliferation and is dose dependent. The half inhibitory concentration (IC50) of Mor-platin is lower than that of cisplatin. Laser confocal images showed that Mor-platin can target cell mito-chondria and enrich cell mitochondria. Transmission electron microscopy images showed that cell mitochondrial morphology changed after Mor-platin treatment. Furthermore, cell mitochondrial membrane is incomplete and mitochondrial cristae are reduced. Cell apoptosis caused by Mor-platin is dose dependent. The three-dimensional tumor spheroid model showed that the cell areas of the group subjected to Mor-platin treatment are smaller than those of the control group. Conclusion:Mor-platin can target cell mitochon-dria, change the cell mitochondrial morphology, inhibit cell proliferation, and thus promote cell apoptosis. It also showed better anti-cancer effects than cisplatin. Furthermore, Mor-platin can inhibit three-dimensional tumor spheroid invasion. These results suggest that Mor-platin is a potential antitumor drug.

Objective In China primary hyperparathyroidism is not a kind of common disease as in the wesyrn countries.This article reports the current status in the diagnosis and treatment of primary hyperparathyroidism in the mainland of China. Methods We collected 730 cages of primary hyperparathyroidism diagnosed and treated in 7 top hospitals for endocrine surgery from 1965 to 2005.Results In this study.652(89.3%)cases were clinically symptomatic while 78(10.7%)cases were asymptomatic:442 cases were positive on 99mTc-MIBI scanning.Bilateral explorations were undertaken in 377 patients and unilateral or uni-gland exploration through the conventional incision in 204 cases.Minimally invasive parathyroidectomy in 143 cases.Endoscopically assisted 2 cm incision was taken in 6 cases for unilateral gland exploration.Pathologically 632(86.6%)cases were identified as adenoma,58(8.3%)cases were of hyperplasia and 40(5.5%)cases were of carcinoma.There were no major postoperative complications.While 20 patients suffering from recurrence or persistent postoperative hyperparathyroidism,the others are of normal or depressed serum level of calcium. Conclusions Preoperative localization is very helpful: Unilateral exploration for parathyroid adenoma is feasible; minimally invasive parathyroidectomy throush minimal incision is a kind of improving procedure for the localized parathyroid adenoma.

Objective To explore the changes and the tendency of breast cancer radiotherapy in China within the past decade.Methods A survey on breast cancer radiotherapy in 1 999 and 2006 was conducted in 4 hospitals located within the southeast coastal region of China,including patients'clinical characteristics,the purpose of radiotherapy and its techniques.Results The percentage of breast conservative treatment(BCT)increased from 3%in 1999 to 13%in 2006.For radiotherapy techniques in 2006 compared with 1999,the growth trend was found in use of breast immobilization devices(80% vs.46%),treatment planning system(70% vs.23%),and CT simulation(14%vs.0).The frequency was increased in the chest wall irradiation after masteetomy(90%vs.67%),but decreased in internal mammary region(30%vs.76%)and axilla(37%vs.69%)treatment.There were no differences in total dose and fractionation prescription.Conclusions BCT was performed more frequently,but postmastectomy radiotherapy was still essential option in China at present.Although some advanced techniques were applied frequently in clinical practice.Simulation,treatment planning,and irradiation target design were urgent for improvement and standardization.

Objective To explore whether chronic forced swimming stress could improve whole brain radiation induced cognitive dysfunction and possible mechanism. Methods Thirty-nine one month old male Sprague-Dawley rats were randomized into sham control group ( C ) , swimming group ( C-S ) , radiation group( R) , and radiation plus swimming group( R-S) . Radiation groups were given a single dose of 20 Gy on whole-brain. Rats in the swimming groups were trained with swimming of 15 min/d, 5 d/w. Rat behavior was performed 3 months after radiation in an order of free activity in an open field and the Morris water maze test including the place navigation and spatial probe tests. Then, the protein expressions of BDNF, P-ERK, T-ERK, P-CREB and T-CREB in the rat hippocampus tissue were assayed by Western blot. Results On the day 2, in the place navigation test of Morris water maze, the latency of swimming group was significantly shorter than that of sham group, the latency of sham group was significantly shorter than that of radiation group, and the latency of radiation swimming group was significantly shorter than that of radiation group(P<0?05). In the open field test, the latencies of the place navigation and spatial probe tests of Morris water maze had no significant difference among four groups (P>0?05). Western blot assay showed that the expressions of BDNF and its downstream signals including P-ERK and P-CREB were markedly reduced by radiation ( P < 0?05 ) , but this reduction was attenuated by the chronic forced swimming stress. Conclusion The chronic forced swimming stress could improve whole brain radiation induced cognitive dysfunction by up-regulating the expressions of BDNF and its downstream signal molecules of P-ERK and P-CREB in hippocampus.

Objective To analyze the correlation factors on bone metastatic rate in prostate cancer patients with prostate specific antigen ( PSA ) less than 20 μg/L.Methods A retrospective analysis was made in 143 prostate cancer patients in Beijing Hospital from March 1997 to December 2013 with PSA less than 20μg/L.Their PSA was 0.05-19.97μg/L and median PSA was 10.72μg/L.According to Gleason score, the patients were divided into three group with Gleason score≤6, 7,≥8.According to T stage, the patients were divided into ≤T2 or ≥T3 .According to PSA, the patients were divided into <4.0 μg/L, 4.0-9.9 μg/L or 10.0 -19.9 μg/L.According to percentage of positive cores on the bone scan, the patients were divided into <33%, 33%-66%or >66%.The correlation of bone scan with PSA, Gleason score, T stage or percentage of positive cores in biopsy was evaluated.Results The bone metastatic rate of each group with Gleason score was 6 ( 9.5%) , 8 ( 16.0%) and 7 ( 23.3%) , respectively.There was significant difference among them ( P=0.025 ) .According to T stage, the positive rate of each group was 6(7.1%) and 15(25.9%), and there was significant difference between them (P=0.036).According to PSA, the positive rate of each group was 2 ( 18.2%) , 9 ( 13.0%) , and 10 ( 15.9%) and there was no significant difference (P=0.153).According to percentage of positive cores on the bone scan, the positive rate of each group was 6 ( 8.8%) , 9 ( 19.1%) and 6 ( 21.4%) and there was no significant difference ( P=0.862).Conclusions In prostate cancer patients with PSA less than 20 μg/L, Gleason score and T stage are significantly correlated with bone metastatic rate.However, bone metastatic rate had no correlation with PSA and percentage of positive cores in biopsy.

Objective To explore the functional mechanism of a Chinese medicine compound “Qiangzhizufang”on rat model of Tourette syndrome ( TS) combined with fear.Methods The rat model of TS combined with fear was established by intraperitoneal injection of 3,3’-iminodipropionitrile (IDPN) combined with acoustic stimulation.After giving different drug lavage treatment, the changes of behavior of the rat models were assessed by field test and behavior test.The content of DA, TH and TH mRNA in the brain tissue was detected by HPLC, immunohistochemistry and RT-PCR, separately.Results Compared with the normal control group, stereotyped behavior and exercise behavior were increased, freezing time prolonged, but the content of DA, TH and TH mRNA in the brain tissue were not obviously changed in the model control group.Compared with the model control group, the stereotyped behavior and exercise behavior were decreased, content of DA, TH and TH mRNA in the brain tissue was decreased in the “Qiangzhizufang” group. Conclusions The Chinese medicine compound“Qiangzhizufang” can improve the behavior in rat models of TS combined with fear.This effect may be realized through down-regurating TH mRNA expression, reducing the content of TH, and reducing the dopamine synthesis.

Objective To identify clinical prevalence of untypical adefovir-resistant mutations of hepatitis B virus (HBV),and to analyze their phenotypic characteristics.Methods 1741 patients with chronic HBV infection were evolved.Untypical adefovir-resistant mutations were analyzed by direct sequencing.Longitudinal analysis was performed by clonal sequencing.Wild-type and mutant HBV genomic amplicons were constructed into pTriEx-HBV 1.1 vector and transfected into HepG2 cells.The replication capacity and the 50％ effective concentration of drugs (EC50) were calculated.Results Patients treated with adefovir alone were more likely to develop rtN238T mutation than those treated with other nucleos(t) ide drugs (x2 =17.10,P ＜ 0.01).The patient received adefovir for 47 months,and then viral rebound and biochemical breakthrough occurred with detection of rtN238T + A181V and rtN238T mutation.Switching-to entecavir therapy suppressed HBV DNA and ALT to an undetectable level and converted all viruses into wild type ones.The reulsts of viral replication capacity showed that rtN238T + A181V strain was higher than rtA181V strain (t =9.54,P ＜ 0.01).Compared to the wild type virus,rtN238T + A181V variant was relatively less susceptible to adefovir.Conclusions rtN238T mutation conferred no resistance to ADV but enhanced natural replication capacity,hence it might represent a novel compensatory drug-resistant mutation for adefovir.