Objective To detect cytokeratin in routine pathology negative regional lymph nodes postoperatively in non-small cell lung carcinoma (NSCLC). To investigate the relationship of lymph node micrometastasis in P-TNM stages NSCLC and survival rates. Methods From Jan. 1996 to Dec. 2003, 107 paraffin-embedded specimens of T1-T4N0-N1M0 NSCLC patients were collected. Anti-cytokeratin(CK) an- tibody AE1/AE3 was applied to detect cytokeratin with Envision~(TM) method in routine pathological negative re- gion lymph nodes in NSCLC, and selected negative control, positive control and blank control. The pulmo- nary hilar lymph node micrometastasis was upward regulated with stage pCK-N1, mediastinal lymph node mi- crometastatsis was upward regulated with stage pCK-N2. The result applied to SPSS11.0 software to process. Results The CK positive rate was 29.9% in all the patients. The CK positive rate was 27% (21/78), 30% (7/23), 67% (4/6)in stage p-Ⅰ, p-Ⅱand p-Ⅲ, respectively. All these data showed the tendency by which detectable rate increased and was accompanied by disease progress. Comparing the annual survival rate and median survival time of the non-micrometastasis group with the mierometastasis group in two groups, the survival rate difference was statistically significant. Comparing the annual survival rate and median sur- vival time in pCK-ⅢA stage with p-Ⅰ-Ⅱstage, pCK-ⅢA stage annual survival rate and median survival time was significantly different (P=0.020). Similarly, comparing the survival rate in pCK-ⅡB stage with p-ⅠB stage, pCK-ⅡB stage survival rate was significantly different(P=0.059). Comparing the survival time of pCK-ⅢA stage with p-Ⅲstage, pCK-ⅡB stage, with p-ⅡB stage, euther survival time difference was statistically significant (P=0.838, 0.518). Conclusions The rate of positive cytokeratin increase is ac- companied by the disease progress in NSCLC. Positive cytokeratin has disadvantagious prognosis. It is showed that pCK-N1 may be equal to p-N1 and pCK-N2 which also may be equal to p-N2. Micrometastasis may affect the UICC staging currently in use.

Objective To observe the influence of coal burning fluorosis on learning and memory ability in rats and reveal its possible mechanisms. Methods Healthy 48 SD rats were divided into control, low-fluoride and high-fluoride group. All rats in fluoride exposed groups were fed with the eom polluted by drying processes with burning coal containing high level of fluoride obtained from the endemic fluorosis area to produce the animal model of fluorosis. The experiment period were 3,6 mouths, respectively. The ability of leaning and memory was measured by Morris test and cholinesterase activity detected by photometric method at 3 or 6 month after experiment, respectively. Results Fluoride contents signifieantlly influenced the escape latency, the numbers of crossing the platforms and the time of staying the platforms(the value of F was 29.29,6.47,6.50, respectively, P<0.01).In addition, the numbers of crossing the platforms and the time of staying the platforms were influenced by the exposed time(the value of F was 16.11,45.59, P<0.01). Furthermore, the fluoride contents and the exposed time had an interaction between the numbers of crossing the platforms and the time of staying the platforms (the value of F was 4.67,5.68, P<0.05 or<0.01). Three months after the experiment, the mean values of escape latency [(14.71± 4.85)s] of rats in highly fluoride exposed group were significantly prolonged as compared with controls [(9.28±4.22)s]; 6 month after the experiment, the mean values of escape latency[(12.42±8.03)s, (17.48± 8.05)s] of rats in both groups exposed to fluoride were significantly prolonged as compared to controls [(7.04± 3.29)s, P<0.05]. The decreased numbers of crossing the platforms[(1.62±0.87)number] and the declined time of staying the platforms[(16.70±5.02)s] were found in the rats exposed to high fluoride as compared to controls [(3.53±1.67 )number, (23.33±5.35)s, P<0.05]. The fluoride contents obviously influenced the activities of acetylcholinesterase and butylcolinesterase (the value of F was 12.83,13.27, P<0.01). On the other hand, the times of breeding also influnced the activities of butylcolinesterase (the value of F was 16.26, P<0.01). In 3 months of the experiment, the activities of butylcolinesterase [(0.55±0.12)kU/g] in low fluoride exposed group were significantly decreased in comparison with controls[(0.73±0.10)kU/g, P<0.05]. The activities of acetylcholinesterase[(0.62±0.42)kU/g] and butylcolioesterase[(0.58±0.10)kU/g] in high fluoride group were significantly decreased as compared to eontrois[(1.41±0.52), (0.73±0.10)kU/g, P<0.05]. The correlation analysis showed that there was a negative correlation between the cholinesterase and the escape latency(r=-0.68, P< 0.01), and a positive correlation between the cholinesterase and the time of staying the platforms(r=0.57, P< 0.01). Conclusions The ability of learning and memory in rats with coal buring fluorosis was decreased, which might be connected to the decreased activity of cholinesterase in a dose-effect correlation.

Objective To compare the efficacies of total gastrectomy (TG) and proximal gastrectomy (PG) for patients with upper gastric cancer.Methods Databases including Medline,Cochrane Library,Web of Science,China National Knowledge Infrastructure,Wanfang database were searched to retrieve literatures on surgical treatment of upper gastric cancer which were published from January 1980 to October 2011.According to different surgical procedures,all the patients were divided into PG group and TG group.Meta analysis were performed by RevMan 5.1.Categorical variables were presented by odds ratio (OR) and 95％ confidence interval (95％CI).Results Thirteen literatures including 2622 patients with upper gastric cancer were retrieved.There were 1464 patients in the TG group and 1158 patients in the PG group.There was no significant difference in the 1-year survival rate between the 2 groups (OR =1.23,P ＞ 0.05).The 3-and 5-year survival rates of patients in the TG group were significantly higher than those of the PG group (OR =1.74,1.45,P ＜ 0.05).There were no significant difference in the 5-year survival rates of patients in TNM Ⅰ,Ⅱ,Ⅳ stages between the 2 groups (OR =0.94,1.31,2.03,P ＞ 0.05),while the 5-year survival rate of patients in TNM Ⅲ stage of TG group was significantly higher than PG group (OR =2.29,P ＜ 0.05) The overall recurrence rate of TG group was slightly lower than that of PG group,with no significant difference OR =0.44,P ＞ 0.05).The local recurrence rate of TG group was significantly lower than that of PG group (OR =0.29,P ＜ 0.05).There was no significant difference in the distal recurrence rate between the 2 groups (OR =0.60,P ＞ 0.05).Conclusions The medium and longterm efficacies of TG are superior than that of PG.The stage of cancer should be taken into account to determine the plan of individual treatment.

To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Drug Delivery Systems ; Erlotinib Hydrochloride ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; Male ; Mutation ; Protein Kinase Inhibitors ; therapeutic use ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; genetics

Objective To investigate the clinical characteristics of the human Adenovirus (HAdv) infections in allogeneic hematopoietic stem cell transplantation ( allo-HSCT) patients and explore the clinical significance of HAdv monitoring .Methods A total of 845 cases underwent allo-HSCT were included retrospectively in Perking University People′s Hospital from October 2012 to August 2014.Peripheral blood HAdv load were monitored twice weekly within 100 days after allo-HSCT, or whenever necessary quantitatively by real-time PCR. Meanwhile, other clinical samples such as stool , urine, and bronchoalveolar lavage fluid ( BLAF ) were also detected qualitatively whenever necessary .The follow-up period was at least six months after allo-HSCT.All clinical data were collected and analyzed .Results The total positive rate of HAdv was 3.4% ( 29/845 ) .The incidence of HAdv infection was higher in children [3.8%(6/155), <18y] than that of adults [3.3%(23/690),≥18y].HAdv infection diagnosed within 100 days after allo-HSCT accounted for 72.4%(21/29) of the total number of positive cases .There were 19 cases detected positive in peripheral blood , 16 cases in stool , 9 cases in urine , and 1 cases in BLAF , respectively.One patient was positive in peripheral blood , stool and urine.The overall median time of HAdv was 69 (13-189) d.The median time was 56 (53 -144) d in stool ,which was earlier than that of in peripheral blood , urine and stool.Among 29 cases of HAdv positive patients , 17 patients were coinfected with Cytomegalovirus(CMV) and 11 casess with Epstein-Barr virus(EBV).Twenty-five cases of HAdv were diagnosed with acute graft-versus-host disease(aGVHD) before HAdv infection, and 4 cases were diagnosed with chronic graft-versus-host disease ( cGVHD ) . The most common clinical manifestation was HAdv enteritis (14 cases), followed by hemorrhagic cystitis (7 cases).Two cases complicated with multiple organ injury ( >2 ) clinically, 1 cases with pneumonia.There were 8 cases of death at the end of follow-up.Conclusions HAdv is an important pathogen causing infection in patients after allo-HSCT. The infenction is characterized with multiple organ involvement .CMV and EBV coinfection is common .HAdv monitoring was of great significance in allo-HSCT patients.

Objective To investigate the effect of Ulinastatin on the delivery of cytokines in patients with septic shock.Methods It was a prospective and controlled clinical study.Seventy-eight patients with septic shock were randomly divided into control group and treatment group and thirty-nine in every group.Patients in treatment group received Ulinastatin 200 000 units intravenous everyday for 3 days,while those in control group received equal volume of normal saline as placebo.At different time points (at 24 th,48 th,72 th hour after start of treatment),the levels of tumor necrosis factor-alpha (TNF-?),interleukin-1 (IL-1),interleukin-6 (IL-6 ),interleukin-8 (IL-8) and superoxide dismutase (SOD) in serum were assayed.Results In comparison with control group,the levels of TNF-?,IL-1,IL-6,IL- 8 of treatment group decreased markedly (P＜0.05,P＜0.01) at different time points,whereas the level of SOD was higher markedly (P＜0.05,P＜0.01) at various time points.Conclusion Ulinastatin has protective effect on patients with septic shock through decreasing the levels of TNF-?,IL-1,IL-6,IL-8 and increasing in the level of SOD.

OBJECTIVETo study the antileukemic activity of L-asparaginase through determining the changes of 4 kinds of amino acids (Asn, Aspa, Glu and Gln) in cell culture medium.

METHODSFollowing L-Asp treatment with designed concentrations and duration, the IC50 (inhibitory concentration 50%) of 8 kinds of common leukemia cell lines (U937, HL-60, Jurkat, NB4, THP-1, Namalwa, Karpass299, K562) were determined by CCK-8 assay. The changes of the 4 kinds of amino acids mentioned above were detected by high performance liquid chromatography (HPLC).

RESULTSThe asparagines in cell culture medium were rapidly exhausted when treated with 0.01 U/mL L-Asp for 4 hrs or 1 U/mL L-Asp for 5 minutes. There were significant differences in the sensitivities to L-Asp of different leukemia cell lines. The sensitivities to L-Asp of various cell lines were dose-dependent. Low concentration of L-Asp resulted in a low IC50 and the IC50 increased following the L-Asp concentration increased.

CONCLUSIONSDifferent leukemia cell lines have different sensitivities to L-Asp, suggesting that exhaustion of asparagines around leukemia cells could not reflect the treatment efficacy of L-Asp. L-Asp antileukemic activity is dose-dependent, which suggests the importance of high-dose L-Asp on childhood acute lymphoblastic leukemia.

Asparaginase ; pharmacology ; Asparagine ; analysis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Humans ; Leukemia ; drug therapy ; metabolism ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy

OBJECTIVETo construct a full-genome hepatitis C virus (HCV) replicon that will allow for direct initiation of replication and generation of infectious viral particles in an in vitro and in vivo cell system.

METHODSSelf-cleaving ribozyme sequences were added to each side of the HCV cDNA clone JFH1 and the replication-deficient clone JFH1/GND, then inserted into the pcDNA3.1 vector downstream of the CMV promoter. The resultant recombinant plasmids, pcDNA3.1-RZ-JFH1 and pcDNA3.1-RZ-JFH1/GND, were tested for activity in vitro and in vivo by transiently transfecting into Huh7.5 cells (5 mug/100 mm culture dish) and injecting by high-pressure tail vein injection into Kunming mice (10 - 30 mug/mouse). Quantitative reverse transcription-PCR, immunofluorescence, immunohistochemistry, and serological testing were performed to determine the replication ability and assess the properties of the recombinant plasmids in the two systems.

RESULTSHCV RNA (1 - 3 * 10(6) copies/ml) was detected in the supernatant of transfected Huh7.5 cells up to 16 weeks after transfection. In addition, the viral particles from the supernatant were able to infect nave Huh7.5 cells. However, only transient viremia was achieved upon tail vein injection of the plasmid, and no HCV antigen-positive cells were detected by immunohistochemistry nor HCV-specific antibodies by serological testing.

CONCLUSIONThe constructed HCV replicon was capable of stable expression in cultured cells and of efficiently generating infectious viral particles in the in vitro system over a long period. However, the HCV replicon did not show infective characteristics in an in vivo mouse system. The full-length HCV replicon may represent a useful tool for in vitro study of HCV pathological mechanisms, possibly including anti-HCV drug screening.

Animals ; Base Sequence ; Cell Line ; Genetic Vectors ; Genome, Viral ; Hepacivirus ; genetics ; physiology ; Humans ; Male ; Mice ; Mice, Inbred Strains ; RNA, Catalytic ; genetics ; Recombination, Genetic ; Replicon ; Virus Replication ; genetics

OBJECTIVETo analyze the relationships between smooth-muscle tumors of gastrointestinal (GI) tract and gastrointestinal stromal tumors (GISTs), and the efficacy of surgical management.

METHODSThe clinical and pathological data of 135 cases of GISTs were collected, including cases of leiomyomas/leiomyosarcoma between 1993 and 2003 and GIST between Jan. 2000 and Jul. 2005. The surgical outcomes were analyzed retrospectively.

RESULTS82.1% of former leiomyomas/leiomyosarcomas was corrected to GISTs. Overall 5-year survival rate was 79.7%. Univariate analysis revealed preoperative metastasis, tumor size, mitotic index, and postoperative metastasis or recurrence were correlated with overall survival in patients with completed resection. Multivariate analysis showed that only postoperative metastasis or recurrence were the indicators of poor prognosis, but without statistical significance (P=0.064). However, multivariate analysis for disease-free survival showed that preoperative metastasis and mitotic index were two independent predictors of poor prognosis (P=0.001 and P<0.001).

CONCLUSIONSMost former leiomyomas/leiomyosarcomas of GI tract should be corrected to the diagnosis of GISTs. Complete surgical resection is the choice of treatment for GISTs. Preoperative metastasis and mitotic index are two independent predictors of poor prognosis.

Female ; Gastrointestinal Stromal Tumors ; diagnosis ; surgery ; Humans ; Male ; Prognosis ; Survival Rate