MB49小鼠膀胱癌腺病毒基因疗法模型的优化

Bladder cancer is regarded as a promising candidate for innovative therapies in the field of immune and gene therapy. In this paper, we present the subcutaneous, metastatic and a novel orthotopic model of murine MB49 bladder cancer in C57BL/6 mice. We further show the potential of using adenoviral vectors together with different transduction enhancers to augment in vivo gene delivery. Finally, we present candidate genes for tumour detection, therapy or targeting. The MB49 tumour grew rapidly in mice. The subcutaneous model allowed for tumour detection within a week and the possibility to monitor growth rate on a day-by-day basis. Injection of MB49 cells intravenously into the tail vein gave rise to lung metastases within 16 days, while instillation of tumour ceils into pretreated bladders led to a survival time of 20 ～ 40 days.     

Studies on the Induction of Cross—Resistance by Low Dose Radiation or by Low Concentrations of Chemicals

The cross-resistance induced by low-level radiation and low concentration of chemical mutagens was studied using human lymphocytes in vitro and mouse bone marrow cells and germ cells in vivo.The chemical mutagens used in these experiments included the MMC H2O2 and CP.The results reported here,in addition to those that have appeared in the literature,show the following reatures documented for the first time:(1)MMC as an adaptive treatment could also induce the adaptive response to radiation-induced cytogenetic damage;(2)esistance against MMC-induced damage could be induced by low dose radiation in vivo;(3)it is in mouse germ cells that low dose radiation or chemical mutagens could induce the cross-adaptation;(4) the cells pretreated by low dose X-rays could show the high resistance to H2O2-induced cytogenetic damage;(5)CP as D1 treat6ment could not induce the adaptive response to radiation and low dose radiation also could not induce the adaptive response to CP.     

Application of Benchmark Dose （BMD） in Estimating Biological Exposure Limit （BEL） to Cadmium

Objective To estimate the biological exposure limit （BEL） using benchmark dose （BMD） based on two sets of data from occupational epidemiology. Methods Cadmium-exposed workers were selected from a cadmium smelting factory and a zinc product factory. Doctors, nurses or shop assistants living in the same area served as a control group. Urinary cadmium （UCd） was used as an exposure biomarker and urinary D2-microgloburin （B2M）, N-acetyl-D-D-glucosaminidase （NAG） and albumin （ALB） as effect biomarkers. All urine parameters were adjusted by urinary creatinine. Software of BMDS （Version 1.3.2, EPA.U.S.A） was used to calculate BMD. Results The cut-off point （abnormal values） was determined based on the upper limit of 95% of effect biomarkers in control group. There was a significant dose response relationship between the effect biomarkers （urinary B2M, NAG9 and ALB） and exposure biomarker （UCd）. BEL value was 5 μg/g creatinine for UB2M as an effect biomarker, consistent with the recommendation of WHO. BEL could be estimated by using the method of BMD. BEL value was 3 μg/g creafinine for UNAG as an effect biomarker. The more sensitive the used biomarker is, the more occupational population will be protected. Conclusion BMD can be used in estimating the biological exposure limit （BEL）. UNAG is a sensitive biomarker for estimating BEL after cadmium exposure.     

A phase II trial of oxaliplatin plus S-1 as a first-line chemotherapy for patients with advanced gastric cancer

Background Palliative chemotherapy has been shown to have a survival benefit for patients with recurrent or metastatic gastric cancer. We conducted a Phase II trial to determine the efficacy and safety of S-1 plus oxaliplatin （SOX regimen） as first-line chemotherapy for patients with unresectable locally advanced or metastatic gastric cancer. Methods Eligible patients had measurable lesions and no previous history of chemotherapy （except adjuvant chemotherapy）. Oxaliplatin was administered intravenously at a dose of 130 mg/m2 on day 1. S-1 was administered orally in doses of 80, 100, or 120 mg/d according to body surface areas of 〈1.25 m2, 1.25-1.5 m2, or 〉1.5 m2 respectively; the total dose was divided into two daily doses on days 1-14. Treatments were repeated every 3 weeks until disease progression or intolerable toxicity occurred. Results Forty-three patients were enrolled in the study. All were assessable for efficacy and adverse events. The objective response and disease control rates were 55.8% and 76.7% respectively. The median follow-up time was 16.5 months. The median progression-free survival time was 7 months （95% CI, 5.8-8.2 months） and the median overall survival time was 16.5 months （95% CI, 9.7-23.3 months）. The one-year survival rate was 54.2%. Major adverse reactions were grade 3/4 neutropenia （9.3%） and thrombocytopenia （20.9%）. Conclusion The SOX regimen with oxaliplatin at a dose of 130 mg/m2 was found to be effective and safe as a first-line chemotherapy in Chinese patients with advanced gastric cancer.     

A Study on Changes of Langerhans Cells in Human Epidermal Cell Culture

Human epidermal cells (ECs) were grown in vitro from normalhuman foreskin and the changes of Langerhans cells (LCs) within theoriginal explants and the epidermal outgrowth were observed in culturewith ATPase staining, HLA - Dr monoclonal antibody indirectimmuno-fluorescence and enzyme labelling avidin-biotin complex (ABC)staining. The capacity of ECs to stimulate the proliferation of allogeniclymhocytes was tested in the mixed skin-lymphocyte culture response(MSLR). The ATPase HLA-Dr positive LCs were gradually decreasedand significantly deformed within the original explants in culture. 70% ofLCs was decreased 3 days after culture. 90% of LCs was lost and the LCsremained in the original explants were only seen their outline 7 days afterculture. LCs approximatively disappeared within the original explants 14to 21 days after culture. The ATPase, HLA - Dr positive LCs have neverbeen found within the epidermal outgrowth. The ECs which were freshlyisolated from skin explants potently stimulated the proliferation ofallogenic lymphocytes in the MSLR. The ECs which were separated fromthe original explants after 3 days of culture were significantly declinedtheir ability to stimulate the proliferation of allogenic lymphocytes in theMSLR. The ECs separated from the original explants after 7 days ofculture were lost their ability to stimulate the proliferation of allogeniclymphocytes in the MSLR. The ECs separated from the epidermaloutgrowth failed to stimulate the proliferation of allogenic lymphocytes inthe MSLR. These results suggested that the ECs grown in our culturesystem were declined their immunogenety of skin allografts.     

Switching from morphine to fentanyl attenuates the decline of μ-opioid receptor expression in periaqueductal gray of rats with morphine tolerance

Background Opioid switching is a therapeutic maneuver to improve analgesic response and/or reduce adverse side effects although the underlying mechanisms remain unknown.The μ-opioid receptor （MOR） has an important role in mediating the actions of morphine and other analgesic agents.This study is aimed at exploring the changes of MOR in the periaqueductal gray （PAG） in rats when morphine is substituted for equianalgesic fentanyl.Methods Forty rats were randomly assigned to five treatment groups：7 days normal saline group （N group）,7 days fentanyl group （F group）,7 days morphine group （M group）,7 days morphine and 7 days fentanyl-switching group （MF group）,and 14 days morphine group （MM group）.Rats repeatedly received subcutaneous injections of morphine sulfate （10 mg/kg） or equianalgesic fentanyl sulfate （0.1 mg/kg） twice daily.Rats＇ antinociceptive response to thermal pain was evaluated by the tail flick latency assay.MOR mRNA and protein expression in the PAG were measured using RT-PCR and Western blotting analyses respectively.Results This study showed that after morphine was substituted with fentanyl on day 8,the tail flick latency （TFL） increased from （3.9±0.4） seconds to （11.4±0.4） seconds.The results also demonstrated that both MOR mRNA and protein expression in the PAG of rats in the MF group were less than that in the M group （P〈0.05） but more than that in MM group （P〈0.05）.Conclusions Equianalgesic fentanyl was still antinociceptive effective in rats with morphine tolerance,which may be due to the switching from morphine to fentanyl attenuating the decline of MOR expression in the PAG of rats.     

Relationship between coronary artery remodeling and cumulative incidence of coronary angiographic lesions with vulnerable characteristics in patients with stable angina pectoris

ackground Development of vulnerable lesions is not limited to the target lesions, but a pan-coronary process. Such lesions are identified by positive remodeling （intravascular ultrasound （IVUS） and complex lesions （angiography））. The prevalence of lesions with vulnerable characteristics in patients with stable angina was not well known. The purpose of the present study was to evaluate the relationship between coronary artery remodeling and incidence of angiographic complex lesions and its calcification in stable angina patients.Methods One hundred and sixty-one stable angina patients （95 males, aged （68±11） years） with 161 de novo target lesions were studied using pre-interventional IVUS. Remodeling index was defined as the lesion divided by reference vessel area; positive remodeling was defined as remodeling index 〉1.05. Besides the 161 target lesions, there were 613 angiographic lesions with 〉30% diameter stenoses, classified as complex or smooth. Multiple complexes were defined as more than one complex lesion in one patient. Stenoses of at least 70% were described as tight. Calcium arc area was used as a new method to quantify coronary calcification.Results Fifty-six patients had positive remodeling target lesion, while 105 did not. The overall number of lesions with a diameter stenoses 〉30% was similar in patients with or without positive remodeling, and the frequency of angiographically complex lesions was higher in positive remodeling patients, especially at non-target site. Calcium arc area was smaller in patients with positive remodeling.Conclusions Positive remodeling on intravascular ultrasound was associated with more complex lesions angiographic findings, especially at non target site. Positive remodeling was found less calcified in patients with stable angina.     

5-aminolaevulinic Acid-photodynamic Therapy for the Treatment of Cervical Condylomata Acuminata

Objective To investigate the efficacy and safety of photodynamic therapy （PDT） with topical 5-aminolaevulinic acid （ALA） on cervical condylomata acuminata. Methods Patients with cervical condylomata （n=30） were allocated into primary and recurrent group, and were given topical ALA under occlusive dressing for 3 hours followed by irradiation with semiconductor laser at a dose of 100 Jcm 2 and a power of 100 roW. The treatment was repeated 7 days later if the lesion was not completely removed after the first treatment. Complete response rate and recurrence rate of wart lesions as well as rate of adverse reaction were analyzed. Results The total complete response rate of PDT was 100% and the total recurrence rate was 5% after 3 months of follow-up. Recurrence rate of recurrent group was significantly lower than that of prior managements （100%, P〈0.01）. The side effects of PDT in patients mainly included mild burning and/or stinging restricted to the illuminated areas, and was significant lower than their own control （25% vs. 100%, P〈0.05）. Conclusion Compared with conventional therapies, topical application of ALA-PDT is a simple, effective, safe, well-tolerated, and low recurrence rate treatment for cervical condylomata acuminata.     

Acetylcholinesterase Characteristics of Termite Queen Exposed to Anticholinesterase Compounds

A regional profile of AChE activity was noted in the Indian termite queen Odontotermes redemanni with the head recording the higher and body the lower range of activity. The enzyme characteristics such as substrate and temperature optima were more or less similar while pH requirement for optimum AChE activity varied from 7.0 to 7.6 In vitro inhibition of head and body AChE was studied using pure and commercial anticholinesterase compounds. Interestingly, the commercial formulations like Metacid-50 and Carbaryl are potent enough at 1×10~(-8) M to produce 50% in vitro inhibition of AChE of head and body regions within 15 min of preincubation. A 20 min of preincubation (t_(0.5)) was necessary to record 50% in vitro inhibition of AChE with known and pure anticholinesterase compounds such as DFP (3.5×10~(-10)M) and physostigmine (3.6×10~(-10)M). It is surmised that (a) the response of the head and body AChE to the commercial formulations of the insecticides may be used as a reliable and sensitive bioindic     

Biokinetics of tritium incorporation into the tissues of rats during continuous ingestion of tritiated water or tritium-labeled food.

Wistar strain male rats were continuously given tritiated water or tritiated wheat as drinking water or food for 70 days. During the ingestion, the tritium incorporation into rat tissues was examined in both wet and dry samples of liver, kidney, testis and blood. The concentration of organically bound tritium (OBT) in dry tissues of rats exposed to tritiated water (HTO) and 3H-food (tritiated wheat) attained an equilibrium within 2-3 weeks after the exposure. The concentration of OBT in dry tissues of rats exposed to HTO also reached an equilibrium within 3-4 weeks after the exposure. However, rats exposed to 3H-food, except for the liver, such an equilibrium state was not reached in other tissues and the OBT concentrations increased gradually throughout the exposure. The relative concentrations of total 3H and OBT at the end of the chronic ingestion of 3H food (70 day), expressed in percentages of the total activity were 1 and 9 times higher than those in rats exposed to HTO, respectively. In both groups, OBT as well as total 3H was almost uniformly distributed among the tissues examined.

     

Treatment of cutaneous T cell lymphoma with low doses of interferon alpha-2b.

OBJECTIVE To evaluate the optimal dose and route of interferon-alpha-2b (INF-alpha-2b) given to the Chinese patients with cutaneous T-cell lymphomas (CTCL).

METHODS Sixteen Chinese patients with CTCL treated with INF-alpha-2b were evaluated. They were divided into groups according to the TNM Classification: I B (8 patients), II A (2), II B (5) and IV B (1). During induction INF-alpha-2b was given intramuscularly every other day or two times a week, beginning with a dose of 1-3 x 10(6) IU. Depending on tolerance, the dosage could be increased weekly to a maximum tolerated dose of 18 x 10(6) IU (mean, 6 x 10(6) IU/week) with administration two or three times a week. Patients with clinical response were given maintenance dosage from the start. Thereafter the dosage was gradually decreased after disappearance of skin lesions. Three patients were treated by combining intramuscular and intralesional injection of INF.

RESULTS Six patients (I B, 2 patients: II A, 2, II B, 2) achieved complete response (CR). Seven patients (I B, 5 patients) showed partial response. The overall response rate in this group was 81.25%. The prominent initial clinical response in three patients was manifested early at one week after intramuscular treatment with INF and then the skin lesions were resistant to the therapy but disappeared rapidly by combining intralesional injection of INF. The conditions of those patients obtaining CR within 3 to 6 weeks were stable for 1-36 weeks.

CONCLUSIONS This study had demonstrated that the overall response rate in treatment of patients with CTCL was 81.25% and higher (85.7%-87.5%) in the subgroups at early stages I and II of the disease. The dosage (mean dose of 6 x 10(6) IU/week) adopted by us is optimal for the Chinese patients with CTCL. Intralesional injection of INF could be recommended.

     

RAT TAIL FLICK ACUPUNCTURE ANALGESIA MODEL

An improved tail flick test, suitahle (or ex- perimental study of acupuncture analgesia k introdueed. It is simpler to conduct and the tail [lick response is better recognized than the con- ventional method. Eleceroacupuncture of bila- teral Zusauli足三里 and Sanyinjiao 三阴交 point cquhralents on the rat resulted in pro- Iongation of the tail flick latency which in- volved descending inhihition from higher centers. The effect of elecLroaCUPunet,ure in this model varied directly within certain limtts wlth the intensity and duration of electra- acupuneture and was fairly stable in a graup of 9 t0 12 0r more rats. The unportance of zroom temperature and constancy of rat tail tem- perature are emphasized.     

Factors influencing ablation of atherosclerotic plaque with argon laser.

The ablative effect of argon laser delivered fibreoptically in vitro on 234 segments of atherosclerotic human aorta was studied. The variables such as energy density, type of atheroma and immersion media were included, and all irradiated specimens were subsequently submitted to histological examination and crater volumes in mm3 to micrometer measurements under light microscope. The results showed 1) a linear relationship between energy dose and crater volume in fibrous atheroma, 2) significantly greater damage or surrounding in the higher energy dose groups, 3) a lower dose response in calcified tissue than in fatty streaks or fibrous atheroma, and 4) immersion of tissue in blood during ablation resulted in a significantly greater dose response than immersion in plasma or saline, and the corresponding surrounding tissue damage was greatest when immersed in blood. Thus, argon laser is both effective and predictable in response during ablating of atheromatous tissue. The efficacy of the process is dependent on immersion medium, while the degree of surrounding tissue damage is dependent on energy dose.

     

Studies on Toxicity,Anti—stress and Hepato—Protective properties of Kombucha Tea

Objective :The objective of the study was to evaluate toxicity,anti-stress activity and hepatoprotective properties of Kombucha tea.Method :Kombucha tea was fed orally for 15 days using three different doses i.e.normal dose,five and ten times the dose,Rats were then sacrficed and various biochemical,and histological parameters were estimated.Anti-stress activity was evaluated either by 1)by exposing animals to cold and hypoxia and estimating the levels of malondialdehyde and reduced glutathione in plasma/blood or 2) by subjecting the animals to restraint stress and recording faecal output.Hepato-toxicity was induced by challenging the animals to an acute dose of paracetamol(1gm/kg)orally and determining the plasma levels of SGPT,SGOT and MDA.REsults:The effect of oral administration of different doses of K-tea to albion rats was examined and the results indicate that K-tea has no significant toxicity as revealed by various biochemical and histopathological parameters.K-tea has been found to prevent lipid peroxidation and fall in reduced glutathione level when rats were exposed to cold and hypoxia in simulated chamber.Further,K-tea has also been found to decrease the Wrap-restraint faecal pellet output in rats.K-tea has also been found to decrease paracetamol induced hepatotoxicity significantly.Conclusion:The study shows that K-tea has anti-stress and hepato-protective activities.     

Acute colonic pseudo-obstruction（Ogilvie′s syndrome）：approach to management

Objective: To assess the value of a parasympathomimetic drug(neostigmine) in the early resolution of acute colonic pseudo-obstruction(Ogilvie′s syndrome).Methods: To study 42 patients in Hawler Teaching Hospital with the diagnosis of acute colonic pseudo-obstruction between years 2004 and 2008.All had abdominal distention and radiographic evidence of colonic dilation,with a cecal diameter of more than 10 cm,and had had no response to at least 36 hours of conservative treatment.We randomly assigned 22 to receive 2.5 mg of neostigmine intravenously and 20 to receive intravenous saline.A physician who was unaware of the patients′ treatment assignments recorded clinical response(defined as prompt evacuation of flatus or stool and a reduction in abdominal distention),abdominal circumference,and measurements of the colon on radiographs.Patients who had no response to the initial injection were received another dose of neostigmine three hours later.Results: Twenty of the 22 patients who received neostigmine had prompt colonic decompression,as compared with none of the 20 patients who received placebo(P<0.001).The median time to response was 5 minutes(range,2 to 30).Fourteen patients in the placebo group and the two patients in the neostigmine group without an initial response received open-label neostigmine;all had colonic decompression.Four patients who had an initial response to neostigmine required colonoscopic decompression for recurrence of colonic distention;two eventually underwent subtotal colectomy.Side effects of neostigmine included abdominal pain,excess salivation,and vomiting.Symptomatic bradycardia developed in four patients and was treated with atropine.Conclusions: In patients with acute colonic pseudo-obstruction who have not had a response to conservative therapy,treatment with neostigmine rapidly decompresses the colon.     

Determinants of Detection of Stones and Calcifications in the Hepatobiliary System on Virtual Nonenhanced Dual-energy CT

Objective To retrospectively determine the features of stones and calcifications in hepatobiliary system on virtual nonenhanced (VNE) dual-energy computed tomography (CT), and to evaluate the possibility of VNE images in diagnosis for those lesions. Methods A total of 128 gall stones and calcifications of the liver found in 110 patients were examined with triple phase abdominal CT scan from July 2007 to December 2011, in which true nonenhanced (TNE) phase and arterial phase were performed with single-energy CT (120 kVp) and portal venous phase was performed with dual-energy CT (100 kVp and 140 kVp). VNE images were generated from the portal venous phase dual-energy CT data sets by using commercially VNC software. The mean CT values for the stone, liver, bile and paraspinal muscle, mean lesion density and size in area dimension, contrast-to-noise ratio (CNR) of lesion to the liver or bile, and image noise were assessed and compared between VNE and TNE images. The effective dose and size-specific dose estimate (SSDE) were also calculated. Results The mean CT values of the lesions measured on VNE images declined significantly compared with those measured on TNE images (164.51±102.13vs. 290.72±197.80 HU,P<0.001), so did the lesion-to-liver CNR (10.80±11.82vs.18.81±17.06,P<0.001) and the lesion-to-bile CNR (17.24±14.41 vs. 21.32±17.31,P<0.001). There was no significant difference in size of lesions area between VNE and TNE images (0.69±0.88vs.0.72±0.85 cm2,P=0.062). Compared to the 128 lesions found in TNE images, VNE images showed the same density in 30 (23.4%) lesions, lighter density in 88 (68.8%) lesions, while failed to show 10 (7.8%) lesions, and showed the same size in 61 (47.7%) lesions and smaller size in 57 (44.5%) lesions. The CT cutoff values of lesion and size were 229.21 HU and 0.15 cm2, respectively. The total effective dose for triple phase scan protocol with TNE images was 19.51±7.03 mSv, and the SSDE was 39.84±11.10 mGy. The effective dose for dual phase scan protocol with VNE images instead of TNE images was 13.29±4.89 mSv, and the SSDE was 27.83±9.99 mGy. Compared with TNE images, the effective dose and SSDE of VNE images were down by 32.05%±3.69 % and 30.63%±2.34 %, respectively. Conclusions Although the CT values and CNR of the lesions decreased in VNE images, the lesions of which attenuation greater than 229.21 HU and size larger than 0.15 cm2could be detected with good reliability and obvious dose reduction. There was good consistency in the size of stones and calcifications in hepatobiliary system between VNE images and TNE images, which ensured the possibility of the clinical application of VNE images.     

Pharmacokinetics of aminophylline delivered to the small intestine and colon using remote controlled capsules

Background A patented remote controlled capsule （RCC） has recently been developed to provide noninvasive drug delivery to selected sites in the human gut that allows assessment of regional gastrointestinal （GI） drug absorption under a normal physiological environment. The objective of this study was to investigate the rate and extent of aminophylline absorption after site-specific delivery of the drug in the GI tract using RCC and a magnetic marker monitoring （MMM） technique. Methods This study was conducted in twelve healthy male subjects, in a three-treatment, randomized, crossover manner with a 7-day washout. Eligible subjects received a 150 mg aminophylline dose through an oral administration, or via a remote controlled capsule, delivered to the small bowel or ascending colon. MMM was employed to monitor the GI transit of the RCC, and the radio-frequency signal was used to activate capsules at target sites. Blood samples were obtained at regular intervals until 24 hours post dose/activation. Plasma theophylline concentrations were measured by a TDx~ System Analyzer. A comparison of the PK profile with the oral dosing route of aminophylline was performed after delivery to the small bowel and colon. Results The RCC was well tolerated in volunteers. The mean capsule activation time for the small bowel and ascending colon was 2.07 hours and 6.08 hours post dose. Aminophylline had similar absorption profiles from the small bowel compared with the stomach, with an area under the curve （AUCt） ratio of 92% vs. the stomach, but a lower absorption profile from the ascending colon, with an AUCt ratio of 47.2% vs. the stomach. Conclusions The proprietary of the RCC and MMM technique offer the opportunity to obtain data on the intestinal absorption of a drug in humans under noninvasive conditions. Aminophylline is rapidly and efficiently absorbed from the small bowel. While colonic absorption was limited by the poor water condition although effective absorption was observed from the ascending colon. This provides an opportunity for rational development of modified-release formulations as well as alternative dosage forms.     

Effects of cadmium on rat sperm motility evaluated with computer assisted sperm analysis

Objective:To study effects of cadmium on rat sperm motility evaluated with computer assisted sperm analysis.Methods:Different doses of cadmium chloride(0.2,0.4,0.8mg Cd/kg BW) were administrated ip to adult male Sprague-Dawley rats.Control animals received the same volume of 0.9% NaCl solution.After 7 days,the rats were sacrificed with their testes removed.A part of one testis was used for testicular sperm head counts and daily sperm production observation.The motility of spermatozoa obtained from cauda epididymides using the “diffusion“ method was measured by computer assisted sperm analysis(CASA).Results:The sperm head Counts and Daily Sperm production decreased significantly in the high dose group.The motility of spermatozoa in the middle ose group was reduced significantly.No motile sperm was found in the high dose group.The results suggest that germinal epithelium was impaired irreversibly in a short time to produce toxic effects on spermatogenesis at high cadmium doses.Conclusion:Cadmium may reduce sperm motility at a dose far below the dose affecting sperm production at this time point.The motiligy of sperm is an early and sensitive endpoint for the assessment of cadmium toxicity on male reproduction.     

A Novel Chitosan CpG Nanoparticle Regulates Cellular and Humoral Immunity of Mice

Objective To develop a safe and novel immunoadjuvant to enhance the immunity and resistance of animals against E. coli infection. Methods An 88-base immunostimulatory oligodeoxynuleotide containing eleven CpG motifs （CpG ODN） was synthesized and amplified by PCR. The chitosan nanoparticle （CNP） was prepared by ion linking method to entrap the CpG ODN that significantly promotes the proliferation of lymphocytes of pig in vitro. Then the CpG- CNP was inoculated into 21-day old Kunming mice, which were orally challenged with virulent K88/K99 E. Coil 35 days after inoculation. Blood was collected from the tail vein of mice on days 0, 7, 14, 21, 28, 35, 42, and 49 after inoculation to detect the changes and content of immunoglobulins, cytokines and immune cells by ELISA, such as IgG, IgA, IgM, IL-2, IL-4, and IL-6. Results The CpG provoked remarkable proliferation of lymphocytes of pig in vitro in comparison with that of control group （P〈0.05）. The inoculation with CpG-CNP significantly raised the content of IgG, IgM, and IgA in the sera of immunized mice （P〈0.05）. The levels of IL-2, 1L-4, and IL-6 in the mice significantly increased in comparison with those in controls （P〈0.05）, so was the number of white blood cells and lymphocytes in immunized mice. The humoral and cellular immunities were significantly enhanced in immunized mice, which resisted the infection of E coli and survived, while the control mice manifested evident symptoms and lesions of infection. Conclusions CpG-CNP can significantly promote cellular and humoral immunity and resistance of mice against E. coli infection, and can be utilized as an effective adjuvant to improve the immunoprotection and resistance of porcine against infectious disease.     

Phase I trial of icotinib, a novel epidermal growth factor receptor tyrosine kinase inhibitor, in Chinese patients with non-small cell lung cancer

Background  The preclinical experiments and studies of congener drugs show icotinib, a new epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, can specifically bind to the tyrosine kinase domain of the EGFR, block the EGFR related signal, thereby inhibit the growth of tumor cell. The objective of this study was to investigate the safety, tolerability and dose-related biologic effects of icotinib in patients with non-small cell lung cancer (NSCLC) in a Chinese patient population.

Methods  This was an open-label, phase I, dose escalation, safety/tolerability trial of oral icotinib (100 to 400 mg), administered twice per day for 28-continuous-day cycles until disease progression or undue toxicity.

Results  Forty patients with stage IIIB (15%) or IV (85%) NSCLC were included in the study. They had mainly adenocarcinoma (85%), with a performance status (PS) of 0 (45%) or 1 (55%) and less than half the patients (45%) had histories of smoking and all were pretreated by at least one regimen of chemotherapy. Patients were assigned to three dose levels of 150 mg b.i.d, 200 mg b.i.d, or 125 mg t.i.d. The follow-up periods ranged from 5 to 80 weeks. Adverse events were found in 35% patients, most of which were mild and reversible. The adverse events mainly occurred in the first 4 weeks and included rash (25%), diarrhea, nausea and abdominal distention. One definite interstitial lung disease (ILD) was found in a patient in the dose of 200 mg b.i.d. According to an 8-week assessment, one (2.5%) patient receiving 150 mg gained complete response (CR) that persisted for 44 weeks, seven (17.50%) patients had partial remission (PR), and 18 (45%) patients had stable disease (SD). The objective response including CR+PR was 20%. The median time of progression-free survival for the 40 patients was 20 weeks (range: 12 to 32 weeks). The response was not affected by pathological type, history of smoking, or numbers of previous therapeutic regimens. No relationship between dose, response, adverse effect, or duration of the study was observed.

Conclusions  Icotinib, given as oral twice daily, showed favorable safety and tolerability. Mild and reversible rash, diarrhea, and nausea were the main adverse events. Antitumor activity was obvious at each dose in heavily pretreated patients. Pharmacodynamic evaluations and further phase II/III trials are in progress.