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 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 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 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 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.

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 curative effect of combined transplantation of bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major.METHODS: Eight thalassemia major patients undergoing combined transplantation of bone marrow and umbilical cord blood of same sibling aged from 4.0 to 7.5 years, 5 boys and 3 girls, were recruited at the Department of Pediatrics, Nanfang Hospital,Southem Medical University from January 2005 to March 2009. The patients were classified into three classes according to Pesarothalassamia classification, class Ⅰ to class Ⅱ 7 cases and class Ⅲ 1 case. Donors ranged 1-4 years received 10 μg/kg per day of subcutaneous granulocyte colony-stimulating factor (G-CSF) for 5 consecutive days. Bone marrow was harvested on the fifth day. Bone marrow and umbilical cord blood of the same sibling then were transfused into the patient.RESULTS: Recovery of hematopoiesis was gained in all patients 4 weeks following transplantation. Seven patients suffered from infection of different degree. Four patients developed mild venous occlusive disease. Two patients developed grade Ⅰ acute graft-versus-host disease (GVHD), and one developed grade Ⅰ chronic GVHD. Seven patients were alive and one died of pulmonary infection and heart failure 32 days following transplantation.CONCLUSION: Combined transplantation of granulocyte colony-stimulating factor primed bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major is safe and effective with promising results. However, complications should be paid high attention following transplantation.

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.

BACKGROUND: According to present theories and our clinical experience, immune ablative and tolerance inducing theory is proposed. Immune ablative means to clear out mutate cell clones and without transfusion of hemopoietic stem cells afterwards; intolerance inducing means to induce animal models not to react to mutate somatic cells, which avoids relapse or new occurrence of autoimmune disease. OBJECTIVE: To explore the effects of immune-ablative and tolerance inducing therapy in treating animal model of immune polymyositis (PM). DESIGN, TIME AND SETTING: Randomized, controlled animal experiment was performed at the Animal Experimental Center of Nanfang Hospital from December 2008 to April 2009. MATERIALS: One New Zealand rabbit, female, weighing 4.1 kg and 36 England guinea pigs, female, weighing 400-500 g, were used. METHODS: New Zealand rabbit's muscle tissue homogenate and complete Freund's adjuvant (CFA) were injected into guinea pigs to make PM animal models. The 28 animal models were randomly divided into intense immune-ablative and tolerance inducing group (Busulfan 1 mg/kg, every 12 hours, totally 8 doses; followed by CTX 40 mg/kg per day for 4 days; then cyclosporine A (CsA) 3 mg/kg per day was given till animals were dead); cyclophosphamide (CTX) group: CTX was given, 10 mg/kg per day for 3days; immune-ablative and tolerance inducing group: Busulfan 0.8 mg/kg, CTX 30 mg/kg, CsA 3 mg/kg; the administration time and dose were the same as group 1. Control group was not treated.MAIN OUTCOME MEASURES: Full blood count (FBC) and biochemical index were tested before and after treatment, and surviving time was recorded. In addition, muscle pathological changes were observed.RESULTS: Compared with control group, number of white cells was significantly decreased in the other groups, and hematopoiesis function gradually restored after administration. The number of white cells in the immune-ablative and tolerance inducing group was the most, and striated muscle pathology showed PM. Following administration, the glutamic oxaloacetic transaminase and creatine kinase of intense immune-ablative and tolerance inducing and immune-ablative and tolerance inducing groups were significantly reduced (P < 0.05, P < 0.01), but no obvious striated muscle pathological changes were found. The glutamic oxaloacetic transaminase, lactic dehydrogenase and creatine kinase in the CTX and control groups remained unchanged. Survival time of intense immune-ablative and tolerance inducing group was the shortest among all groups, and there was no significant difference between CTX and control groups. The animals in immune-ablative and tolerance inducing group survived for the longest time. CONCLUSION: Immune-ablative and tolerance inducing therapy has preferable effect on treating animal models of PM, and its prognosis is better than intense immune-ablative and tolerance inducing therapy and regular CTX therapy.