Objective To measure the serum levels of endostatin in lung cancer patients in different stages and with histological types, and explore the clinical value of serum level in early diagnosis of lung cancer.Methods ELISA was used to detect the level of serum endostatin in 81 patients with lung cancer (lung cancer group), 23 patients with benign lung disease (benign lung disease group), 20 healthy controls (healthy control group). The serum levels of endostatin were analyzed in lung cancer patients in different stages and with histological types.Results The serum level of endostatin in lung cancer group was (2.53±8.75) ng/mL, significantly higher than that in benign lung disease group (4.63±1.12) ng/mL and healthy control group (4.53±1.24) ng/mL. In lung cancer group, the serum level of endostatin of stage Ⅰsubgroup and Ⅱsubgroup was significantly higher than that of stage Ⅲ subgroup. The differences were statistically significant. There were statistical differences of serum level of endostatin among the lung cancer patients with different histological types.Conclusion Serum endostatin might be used as an indicator of early diagnosis of lung cancer.

Objective To identify the spectrum of pathogenic microorganisms of urinary tract infection and the drug resistance in a hospital setting. Methods The pathogenic microorganisms isolated from 3117 mid-stream urine samples of patients admitted in Hangzhou First People's Hospital from 2005 to 2007 and their drug resistance results were retrospectively analyzed. Results Bacteria were the most prevalent microorganisms in the urinary tract infection, and followed by fungus, mycoplasma and chlamydia. Escherichia eoli accounted for the largest proportion of gram-negative bacteria, in which the ESBLs positive strains accounted for 51.2%, and their drug resistance rate was much higher than that of ESBLs negative strains. Main gram-positive coccobacteria was all sensitive to vancomycin, and relatively sensitive to nitrofurantnin and ampicillin. Conclusions Escherichia coli continue to prevail upon the spectrum of pathogenic microorganism of the urinary tract infection, and the fungus, mycoplasma and chlamydia infections are rising. Antibacterial agents should be used under the guidance of drug sensitivity test, and the combined use should be avoidd.

Objective To explore the invasion effect of CXCR3 overexpression on T lymphoblastic leukemia (Jurkat cells) with chemokine receptors. Methods Mouse CXCR3 was amplified by RT-PCR and overexpressing CXCR3 lentivirus carrying GFP&Puromycin (puro) was constructed. CXCR3 expression on infected Jurkat cells surface was detected by FCM. Constructed cells were seeded in Transwell invasion model to study whether CXCR3 overexpression would increase the invasion or not. Results GFP expression on Jurkat cells was less than 10% after 96 h lentivirus infection. CXCR3 expression was 90% higher than vector group , and GFP expression reached 90% after screening. Therefore, Jurkat cells with stable overexpression of CXCR3 were successfully constructed. Invasion rate of Jurkat CXCR3 cells was [(12.71 ± 1.03)%], which was significant higher than that of vector control group [(6.82 ± 0.49)%], (P < 0.0001). Conclusions CXCR3 expression on leukemia cells is closely associated with leukemia invasion. The increase of CXCR3 expression can enhance the invasion of leukemia cells, and may be one of the mechanisms of T lymphoblastic leukemia invasion.

Objective To observe the effects of hyperbaric oxygen therapy(HBOT) on Glasgow coma scale(GCS) in patients with traumatic brain injury(TBI) and the influences of course and initiating time of HBOT on the therapeutic effects.Methods 105 cases of TBI patients,which performed HBOT more than 30 days in HBOT Center of Southwest Hospital,were analyzed retrospectively.The GCS improvements were compared with 29 cases of TBI patients without HBOT during the same period.They were also compared between patients with different severity,initiating times and courses of HBOT.Results The GCS improvement of patients with HBOT was 3.97?2.65,especially in severe TBI patients(5.22?2.49),Both were higher than that without HBOT(2.38?2.16)(P

Objective:To clone and express human Fas activation domain(FasAD)with the bioactivity.Methods:The FasAD cDNA was amplified by seminested RT PCR,and then inserted into the prokaryote vector of pTYB12 expressed intein protein to construct the recombinant plasmid of FasAD pTYB12.The FasAD peptide was expressed and purified in IMPACT TM CN system by the method of one step affinity purification.Results:Sequence analysis revealed that cloned FasAD cDNA sequence was completely same as that of Genebank record(M67454).Soluble fusion protein was successfully expressed by induction of IPTG.The FasAD peptide with a molecular weight of 5 000 was obtained by purification and was recognized by the rabbit anti human Fas polyclonal antibody in Western blot analysis.It’s activity of inhibition of apoptosis induced by rhFasL can each to 70% in primary biological detection.Conclusion:The above results indicated that FasAD peptide could be prepared by using the IMPACT TM CN system,thus laid a relatively experimental foundation for further research of relationship between structure, function and interaction with it’s ligands,and for further development of biological immune modulator. [

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Objective To investigate the autophagy and apoptosis in acute myelogenous leukemia U937 cell induced by Sirolimus.Methods U937 cells were subcultured, and blank control group(normal) and Sirolimus treated groups(12 h, 24 h,48 h) were established.The Sirolimus treated groups were treated by 2 μmol/L concentration of Sirolimus for 12 h,24 h and 48 h, respectively.The cell morphology of U937 cells treated by Sirolimus was observed after 12 h,24 h and 48 h.The survival rate of cells was detected by cell counting kit-8 method.Cell apoptosis was detected by flow cytometry using Annexin V-FITC/PI double labeled.Real-time PCR was used to detect the level of mRNA expression in autophagy specific protein maker mictotubule-associated protein light chain 3 (LC3)-Ⅱ in different treated times by Sirolimus.Sirolimus LC3 protein expression levels after treatment were detected by Western blot method.Results Under inverted microscope, the cell number of Sirolimus treatment group reduced gradually after 12 h ,24 h and 48 h culture, volume of cells became smaller, cells got ruptured, and the nucleus pycnosis and cellular debris increased.With the extension of time, U937 cells survival rate was falling, and there was statistical differences compared with those of the control group(P =0.031).With Sirolimus treatment, U937 cells after 12 h,24 h and 48 h, U937 cell apoptosis rate increased, and there were statistically significances, compared with those of the control group (P =0.027).With Sirolimus treatment U937 cells after 12 h,24 h and 48 h,LC3-Ⅱ mRNA expression and protein expression were down-regulated compared with those of the control group, and there were statistically significances (P =0.029).Conclusions Sirolimus can induce autophagy and apoptosis in U937 cells.Autophagy protein LC3-Ⅱ in gene and protein expression levels were lowered, and LC3-Ⅱ may play an important role in regulating the leukemia cell autophagy.

BACKGROUND: Directly percutaneous injection of protein-denaturant hydrochloric acid (PDHA) into tumors can lead to fast killing of tumor, sustained drug release and prevention of in situ recurrence of tumor. However, whether implants can be used combined with denaturant still remains unknown. OBJECTIVE: To investigate the compatibility of fluorouracil implants and PDHA (6 mol/L). DESIGN, TIME AND SETTING: Observational study was performed in the Hefei Industry University between October 2006 and March 2007. MATERIALS: A total of 78 Wistar rats, weighing (200i20) g, half males and half females, were used for testing drug release in vivo. Drugs fluorouracil implants (H20030345; columniform particle, diameter 0.8 mm, length 4 mm; specifications: Fluorouracil 2 mg/particle; batch number: 20060922; meeting the National Drug Quality Standards [WS1-(X-103)-2005Z]) were provided by Wuhu Zhongren Pharmaceutical Company,Ltd. Hydrochloric acid (37%) was analytical reagent. METHODS: 96 tubes of the implants and PDHA were kept at (37.0± 0.5) ℃. Each time, six samples were collected at 1, 8, 16, 24, 96, 120, 168, 240, 360, 432, 480, 528, 600, 720, and 960 hours after incubation. Appearance of the implants was observed by microscope. Stability of fluorouracil in PDHA was determined by HPLC and ultraviolet absorb method. Based on the entering quantity and residual quantity of fluorouracil, the release rates were calculated. MAIN OUTCOME MEASURES: The approximate solubility, stability and morphological change of fluorouracil in denaturant and the corresponding drug release character in both denaturant and rats in vivo. RESULTS: At (37.0±0,5) ℃, the fluorouracil was stable for 960 hours in PDHA, the saturated concentration of fluorouracil was (22.72±0.04) g/L. The appearance of implants was intact. The surface was porous. Compared with the speed of releasing drug in rats, the speed of releasing drug was faster in the early stage of release process and slower in the later stage. The drug release was incomplete. At 1, 24, 96, 360 and 960 hours, the implants' release rates were (11.9±6.7)%, (37.9±5.3)%, (52.6±4.5)%, (75.3±3.8)%, and (85.5±2.1)%, respectively. CONCLUSION: The fluorouracil implants and hydrochloric acid (6 mol/L) are compatible and no influence is detected during the observation.

AIM: To assess the effect of RNAi on suppressing the exogenous reporter gene expression in mammalian neurons,and explore the effect of siRNA quantitation on interference efficiency.METHODS: Exogenous green fluorescent protein(GFP) expression vector was transferred into neuro-2a cells,and then the small interference RNA targeting GFP mRNA(siGFP) synthesized by transcription in vitro at three different concentration was used in this experiment.RESULTS: The results showed that the neuro-2a cells can be transfected efficiently and siGFP can inhibit GFP expression greatly.CONCLUSION: RNAi can be applied into mammalian neurons successfully.The research on siRNA quantitation will provide technique support for studying the gene function of neurons in the future.

Objective To investigate the efficacy and safety of fluid resuscitation with 75 g/L NaCl solution in traumatic hypovolemic shock using Meta-analysis. Methods Based on the inclusion criteria,the randomized controlled trials (RCTs) were identified by retrieving the databases including PUBMED,EMBASE,OVID,Cochrane library and EBSCO.The quality of studies was also evaluated.Indices including systolic blood pressure,hemoglobin,serum Na level and mortality were extracted from the enrolled RTCs and were comparatively analyzed between the 75 g/L NaCl group and the isotonic saline group for evaluating the safety and efficacy of 75 g/L NaCI solution.The extracted data were analyzed by using the Review Manager 5.0.25. Results Six RCTs were eligible for Meta-analysis.The 75 g/L NaCl group displayed remarkable increase in systolic blood pressure and decrease in hemoglobin level,compared with the isotonic saline group ( MD =6.23,95％ CI 2.78-9.69,P ＜ 0.01 ; MD =-6.11,95％CI -8.25-3.96,P ＜ 0.01,respectively).Short-term hyperosmolar state in 75 g/L NaCl group was increased significantly but was considered acceptable ( MD =7.97,95％ CI 7.55-8.38,P ＜ 0.01 ).No significant difference in mortality was found between 75 g/L NaCl group and isotonic saline group ( RR =0.96,95％ CI 0.84-1.10,P ＞ 0.05). Conclusion Fluid resuscitation with 75 g/L NaCl solution is safe and effective for traumatic hypovolemic shock.