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Objective@#To develop a method for rapid detection of influenza virus and its subtypes based on flow fluorescence technology, and then detect the clinical specimens by our established method .@*Methods@#We designed degenerate primers and specific probes, synthesized plasmids, used Luminex platform to establish detection method and later detected 430 clinical specimens, and compared the result with those of Fujian Center for Disease Control and prevention.@*Results@#A method for the simultaneous determination of influenza viruses A, B, C and its subtypes (H116, N19) was established. The time consumption was 3.5 hours, with good specificity, high sensitivity and feasible stability. The detection result of 430 clinical specimens showed high consistency with the result of Fujian Center for Disease Control and Prevention.@*Conclusions@#We established a method for simultaneous determination of influenza viruses and its subtypes, high sensitivity, specificity and stability.
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Objective To analyze the clinical manifestations of pulmonary fungal disease in patients with different pathogen infection and different immune states. Methods A retrospective analysis was carried out with the clinical data of 78 patients with pulmonary fungal disease who were treated in Fujian Provincial Hospital from June 2012 to June 2016. Results The identified pathogens included Cryptococcus (78.2％, 61/78), Aspergillus(17.9％, 14/78), Talaromyces marneffei, Mucor, and Trichosporon asahii (one each). Cryptococcus was mostly found in healthy individuals without underlying disease and Aspergillus largely infected those with underlying disease. Diabetes mellitus was the most common underlying disease. Pulmonary fungal infection was confirmed by pneumonectomy (41.0％, 32/78), CT-guided percutaneous transthoracic biopsy (43.6％, 34/78), transbronchial lung biopsy (14.1％, 11/78), or blood culture (1.3％, 1/78). Pulmonary cryptococcosis was often asymptomatic (47.5％, 29/61). Hemoptysis was only found in the patients with underlying disease. The patients with pulmonary aspergillosis showed higher prevalence of hemoptysis (57.1％, 8/14) than the patients with other pulmonary fungal diseases. Bronchoscopy usually gave negative finding in case of pulmonary cryptococcosis (77.8％, 14/18). Inflammatory exudation was the primary finding of pulmonary aspergillosis (6/10). Intraluminal necrosis and neoplasia were only found in the aspergillosis patients with underlying disease. The primary imaging findings in pulmonary cryptococcosis was nodule or mass (78.7％, 48/61). Halo sign and crescent sign were rarely found in pulmonary aspergillosis. Of the 78 patients, 45 (57.7％) patients received pharmacological therapy alone, 15 (19.2％) surgical treatment alone, and 18 (23.1％) received drugs in combination with surgery. Six patients died, 25 lost to follow-up, and 47 with stable disease. Conclusions The clinical characteristics of pulmonary fungal disease vary with the pathogen and the immune states of patients. Clinical symptoms and immune status of patients should be taken into account when making diagnosis of pulmonary fungal disease for the purpose to speculate the probable fungal pathogen and choose the most appropriate diagnostic tool.
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Objective To establish a mouse model of lung adenocarcinoma brain metastasis with human luc+-PC?9 cells stably expressing luciferase and to compare the evaluation values of bioluminescence imaging and18 F?FDG ( 18 F?flu?orodeoxyglucose) SPECT/CT in these models. Methods Suspension of luc+?PC?9 cells was injected into the left ventri?cle of BALB/c nude mice to establish a mouse model of brain metastasis from lung cancer. Bioluminescence imaging and18 F?FDG SPECT/CT were used to evaluate the metastasis of tumors as compared with HE?staining pathology as a golden standard. Results The success rate of brain metastases was 85% through injecting luc+?PC?9 cells into the left ventricle. The number of tumor cells was positively related to the intensity of light, with a linear correlation (R2 =0. 96). Fluores?cence was observed in the brain, spine and femur by bioluminescence imaging, and the metastases were confirmed by H&E pathological examination. 18 F?FDG SPECT/CT observed abnormal density collective foci in the spine or femur but not in the brain. Conclusions Injection of tumor cell suspension into the mouse left ventricle is a good method to establish a brain metastasis of lung cancer. Bioluminescence has a higher sensitivity and specificity in detecting brain metastasis and bone metastasis, with advantages of real?time, dynamical and non?invasive detection of tumor metastasis growth. 18 F?FDG SPECT/CT does not have superiority in detection of brain metastases but is suitable for detecting bone metastasis.
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Objective Previous studies regarding the effects of tamoxifen ( TAM) on the thin endometrium are rare .The aim of this study was to explore the effects of TAM on patients with thin endometrium undergoing frozen thawed embryo transfer ( FET ) . Methods One hundred and thirty three patients with thin endometri-um undergoing FET treatment were recruited from January 2014 to June 2016, who canceled embryo transfer ( ET) or after FET due to thin endometrium in natural cycle or hormone replacement therapy cycle .Patients were randomly divided into letrozole ( LE,n=72) group or tamoxifen (TAM,n=61) group.All of the patients started to have oral pills of Estradiol Valerate 4 mg/d on the third day of menstruating cycles , then 6mg/d on the eighth day ,after 10~12 days then having ultrasonic monitoring of endometrial thickness and blood estradiol (E2), progesterone levels, It′s called endometrial preparation for hormone replacement cycle .To letrozole, tamoxifen group,the way of endometrial preparation were as follows:patients started to have oral pills of LE 2.5mg/d,TAM 40 mg/d on the third day of menstruating cycles for 5 days, then having ultrasonic monitoring and used drug of human chorionic gonadotropic hormone ,It′s called HCG day .After the dominant follicle ovulation then took progesterone intramuscular injection 40 mg/d, oral progesterone 20 mg/d to change endometrium ,then to transplant cleavage embryos or blastocysts after taking 3 or 5 days of progesterone , It′s called embryo transplanting day .The way of TAM endometrium preparation was called TAM cycle .The general data , hormone levels and clinical out-come between two groups were analyzed . Results The serum estradiol level of LE group both on HCG and transfer day [(1193.80± 629.64)ng/L vs (2776.30±157.34)ng/L;(1195.90±820.30)ng/L vs (2129.40±1208.71) ng/L,P=0.000] were statistically lower, serum luteinizing hormone level were statistically higher than TAM group [(20.48±15.50)IU/L vs (10.59±8.34)IU/L,P<0.05];im-plantation rate of LE group were statistically lower than TAM (39.32%vs 45.83%,P=0.001).The endometrial thickness and serum E 2 and P levels in TAM cycles were significantly higher compared with those in hormone replacement therapy cycle [(8.49±1.36)mm vs (6.43±0.96)mm,P=0.018]. Conclusion Tam compared LE with patients of thin endometrium undergoing FET can increased en -dometrial thickness and improve implantation rate ,thus Providing a new solution to thin endometrium .
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Objective To observe the ultrastructure of blood-brain barrier in the nude mouse model of brain me-tastases from lung cancer by transmission electron microscopy using lanthanum nitrate tracing.Methods PC-9 cells (1 × 106/0.1 mL) in logarithmic phase were respectively injected into six nude mice ( model group) selected from eight nude mice randomly via the left ventricle, the other two mice without any treatment as the control group.The general status of the mice was observed after implantation.In the fourth week all the mice were sacrificed and brain tissue samples were taken and prepared for transmission electron microscopic observation using lanthanum nitrate tracing.besides, the lung and brain were removed and stained with HE to detect the presence of tumor metastasis.Results Mice in the model group began to lose weight almost simultaneously in the third week and became moribund slowly, and were all sacrificed at the fourth week when showing clear signs of cachexia.At autopsy, the thoraxes were clear, with normal lungs.Histology showed evidence of brain metastasis in all the six mice.The electron microscopy showed that lathanum nitrate tracer was escaped from the capillaries and diffusely or sparsely distributed in the brain tissues of the model group mice, however lathanum nitrate tracer was still confined in the capillary lumen in the mice of control group.Conclusions The diffuse lathanum nitrate tracer in the brain parenchymal tissue indicates the impairment of blood-brain barrier in the nude mouse model of lung cancer brain metastasis and the formation of these metastases is accompanied with the destruction of blood brain barrier.
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By expounding the definition and application status quo of egg freezing with non-medical factors, this paper ethically analyzed the conflict between egg freezing with non-medical factors and assisted reproductive technology standard management. After systematic evaluation on effectiveness, safety, economy and ethics, it sug-gested that the egg freezing with non-medical factors required careful consideration.
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Objective To establish an appropriate animal model of brain metastases from lung cancer in nude mice by thoracic orthotopic implantation in the chest or left ventricular injection , and to serve further studies on the mechanisms of lung cancer brain metastasis .Methods PC-9 cells (1 ×106/0.1 mL) in logarithmic phase were respectively injected into 18 nude mice by orthotopic implantation in the chest or left ventricular injection ( n=9 each group ) .The statuses of nude mice were observed after implantation .Animals showing clear signs of dyscrasia were killed .At autopsy, the lung, brain, liver and kidney were removed and histological sections were stained with H /E to detect the presence of tumor cells . Results In the thoracic orthotopic implantation group , three weeks after implantation , the number 4, 6, 9 mice showed tumor nodules in the chest wall , they began to lose weight in the fourth to sixth week differently , showing signs of dyscrasia gradually , and were sacrificed at the fifth to seventh week .The thoracotomy revealed that the whole thorax was occupied by many large lung cancer masses , spreading into bilateral ribs , pleura and spinal vertebra , with scarce eroded , compressed , pale and distorted lung tissues left .Histological examination with HE staining showed the presence of neoplasms in their lung tissues but only the number 6 mouse showed metastatic lesions in the brain tissue .In the left ventricular injection group, the mice almost began to lose weight in the third week simultaneously and became moribund slowly , which were all sacrificed at the fourth week .After thoracotomy , the thoraxes were clear except the number 11 and 18 mice which appeared 2-3 tiny tumor foci in the chest wall , with normal lung tissues .Histological examination with HE staining showed the pres-ence of brain metastases in all the nine mice .The rate of brain metastases from lung cancer in the left ventricular injection group was 100%, compared with 11.1% in the thoracic orthotopic implantation group .Conclusions The establishment method of mouse model by left ventricular injection shows significantly higher rate of lung cancer brain metastases than that by thoracic orthotopic implantation .
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Objective To establish an optimal animal model of pulmonary metastasis of human lung adenocarcino-ma, to serve further investigation of mechanism of lung adenocarcinoma metastasis.Methods Eleven nude mice aged 4-6 weeks were used in this study.Suspension of human lung adnocarcinoma A549 cells (0.1 mL, 107 cells/mL) was injec-ted into the tail vein in nude mice.From four weeks after inoculation, two nude mice were killed each time at 4, 5, 6 weeks after the tumor cell injection at random for examination.The remaining 3 mice were killed at the end of the experi-ment.At autopsy, the lung, brain, liver, kidney and other organs were removed, fixed in neutral buffered formalin and embedded in paraffin.Sections were cut and stained with hematoxylin-eosin, and examined by histopathology.The number of metastatic foci was counted.Results No mouse died after tumor cell inoculation.Serially euthanized mice revealed evi-dence of gradually increasing pulmonary metastases in the mice:No metastasis was found before 4 weeks after tumor cell in-oculation, the first histological metastases appeared at 5 weeks, gross metastatic foci were observed at 6 weeks, widely spread metastatic foci were observed at 7 weeks, and the remain 3 mice developed cachesia at 11, 13, and 14 weeks after tumor cell inoculation.Mediastinal lymph node metastases were found in the nude mice by 11 weeks after tumor cell inocu-lation.Conclusions We have successfully established a nude mouse model of pulmonary metastasis by injecting human lung adenocarcinoma A549 cells into the tail vein.
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Objective To screen the anti-tumor prescription (Shengzao Fang) of Shengma (Rhizoma Cimicifugae) combined with Zao Jiao-ci (Spina Gleditsiae), and evaluate its anti-tumor effects. Methods The lung metastatic model of 4T1 breast cancer and the 4T1 subcutaneous transplanted model were established, both models of mice were randomly divided into model group, Shengma group, Zao Jiao-ci group, Shengzao Fang first group, Shengzao Fang second group and Shengzao Fang third group, 10 mice for each group. The proportion of Shengzao Fang was ascertained by their anti-tumor effects. MTT assay was used to investigate the effects of Shengzao Fang on 4T1 breast cancer. The concentration of type 1 collagen (Col-Ⅰ), platelet-activating factor (PAF), tissue inhibitor of metalloproteinase 1 (TIMP-1), tyrosine kinase receptor A (TrkA) and tyrosine kinase receptor B (TrkB) in serum from tumor-bearing mice was tested by enzyme-linked immunosorbent assay (ELISA). The effects of Shengzao Fang on matrix metalloproteinase (MMPs) was examined by Gelatin matrix method. Results Compared with the model group, the prescription of Shengma∶Zao Jiao-Ci=2∶1 significantly suppressed lung metastasis of 4T1 breast cancer, showing fewer lung nodes, lower lung metastasis rate and highest tumor inhibitory rate to 4T1 subcutaneous transplanted model. Although the concentration of serum Col-Ⅰ, PAF, TIMP-1, TrkA and TrkB was decreased in all treated group, the prescription of Shengma∶Zao Jiao-ci=2∶1 had the strongest activity, and its inhibitory effect on the expression of MMP-2 and MMP-9 was strongest, too. Conclusion The prescriptions of Shengma combined with Zao Jiao-ci had different antitumor activity, the strongest activity was exhibited when shengma∶Zao Jiao-ci was 2∶1, suggesting that 2∶1 was optimization for Shengma combined with Zao Jiao-ci.
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ObjectiveTo investigate the efficacy of highly active antiretroviral therapy (HAART) for HIV/HCV co-infection patients. MethodsA randomized and double blinded trial was conducted in sixty-three HIV/HCV co-infected patients ( group A) and 62 HIV infected patients ( group B). The group A (study group) was further divided into A1, A2, A3 subgroups randomly by Spw-Pb network data system, and were given three different HAART regimens based on nevirapine (NVP), efavirenz (EFV) and lopinavir/ritonavir(LPV/r), respectively. CD4+ T lymphocyte counts, HIV virus load, glutamate-pyruvate transaminase (ALT) were detected at baseline and at the endpoint of study (48 weeks). SPSS 13.0 was used for statistical analysis. One-way ANOVA and LSD-t tests were performed. ResultsAfter 48 weeks treatment, HIV RNA became negative in 59 patients of group A (59/63, 93.7％ ), while that in group B was 61 (61/62, 98.4％ ) (x2 =0. 159, P ＞ 0.05 ). CD4+ T lymphocyte count in group A was (208 ± 77 )/μL, which was significantly lower than that in group B (263 ± 78)/μL (t =-2. 759, P = 0. 008 ).ALT level in group A was (57 ±49)U/L, which was significantly higher than in group B (31 ± 14) U/L (t = 2. 027, P = 0.047). CD4 + T lymphocyte count in group A3 was significantly higher than that in A1 (t=-2. 191, P =0.045), while ALT level in A1 was much higher than that in subgroups A2 and A3 ( t = 2.568 and 2.478, P ＜ 0. 05 ). The incurrence of drug-induced hepatitis in HIV/HCV co-infected group was much higher than that in HIV infected group (55.5％ vs. 27.4％, x2 = 10. 182, P = 0.001 ).ConclusionsHCV co-infection in HIV patients shows no impact on virological response to HAART, but the immunological response is poorer.Hepatotoxicity is common among patients receiving HAART, especially those who are receiving NVP containing regimens. LPV/r based regimens are recommend for HIV/HCV coinfected patients.
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<p><b>BACKGROUND</b>Livin is a novel inhibitor of apoptosis protein(IAP),recent studies showed that it overexpressed in many carcinomas including lung cancer and contributed much to the cancerous development.The objective of this study is to explore the expression of the two isoforms of livin in lung cancer tissues and their relationship with histological types and chemotherapy,and to explore their relationship to the expression of caspase-3 as well.</p><p><b>METHODS</b>Expression of livin α,livin β and caspase-3 mRNAs were detected by reverse transcription polymerase chain reaction(RT-PCR) assay in lung cancer tissues as well as in controls.</p><p><b>RESULTS</b>Livin α and livin β were expressed in 12 of 27 and 19 of 27 lung cancer tissues respectively,much higher than those in lung para-cancerous(0/6,0/6) or benign disease lung tissues(0/12,1/12)(both P < 0.01).Moreover,the positive rate was 7/14 and 9/14 in lung adenocarcinoma and 4/12 and 9/12 in squamous and large cell carcinoma respectively,and both of them were detected in one small cell carcinoma.The levels of these two isoforms in lung cancer were significantly higher than those in controls by Gel Imaging System(both P < 0.05),the level of livin α was remarkably higher in adenocarcinoma than that in squamous cell carcinoma(P < 0.05),while the level of livin β was similar in both carcinomas(P > 0.05).Meanwhile,the level of caspase-3 in lung cancer was significantly lower than that in controls,the levels of either each of two isoforms or their sum were negatively associated with that of caspase-3(P < 0.05,P < 0.01,P < 0.01).Two isoforms of livin mRNA expression seemed to increase after chemotherapy but not related to clinical stages(P > 0.05).</p><p><b>CONCLUSIONS</b>Two isoforms of livin are differently expressed in different histological types of lung cancer and may contribute to corresponding cancerous development;the levels of livin are negatively associated with those of caspase-3,this may due to the fact that livin could resist against apoptosis;high expression of livin seems to be related to chemotherapy but not clinical stage.</p>
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Objective To evaluate the clinical application of helical CT 3D reconstruction technique in the dental orthopaedics. Methods The helical CT was performed with 3.0 mm slice thickness and 1.0 pitch in 41 patients with dental orthopaedics. The 3D reconstructions, including maximum intensity projection (MIP), surface shaded display (SSD), and multiplanar reconstructions (MPR), were made for all the cases. Results Thirty-seven of the 41 patients showed malalignment, tilt, rotation, overlap of the teeth and the different space between the longitudinal axes of the teeth. Twenty-five cases of them have shown 36 buried teeth in all. The axial images covered all the information. SSD demonstrated the external contours and entire morphologies of the teeth and the mandible with the relationship of the teeth alignment and the mandible. MIP clearly manifested the full view and the longitudinal alignment of the teeth. Among the 36 buried teeth, there were 29 palatally and 7 labially presented teeth, and they were morphologically delineated on MIP through various angles. Conclusion The helical CT 3D reconstruction is a new technique to display the stereoscopic configuration of teeth. The combination of axial images and MIP, SSD, and MPR provides valuable anatomic and diagnostic information helpful for the surgeons to structure and determine the treatment protocol for the dental orthopaedics.