Objective:To study the effects of human CBS(Cord Blood Serum) on hematopoietic cells culture in vitro.Methods:CBS and/or different combinations of cytokines were added to the culture system derived from human bone marrow mononuclear cells.Colony forming capacity of colony forming unit-granulocyte/ monocyte (CFU-GM)?colony forming unit-granulocyte/ erythrocyte/ megakaryocyte/ monocyte (CFU-GEMM)?cobble-stone area forming cells (CAFC)?long-term culture initiating cells (LTC-IC) were observed.Results:CBS had certain cytokine-like stimulating activity that could promote the proliferation and differentiation of hematopoietic cells of bone marrow in vitro. By comparison with cytokines, its GM-CSF-like activity was equal to 65.6 ?g/L rhGM-CSF, and its IL-3-like activity was equal to 2.9 ?g/L rhIL-3.Conclusion:CBS contains stimulating activities for proliferating and differentiating of hematopoietic cells. CBS alone can maintain hematopoietic cells culture in vitro. This study showed the prospect of clinical application of CBS.

Objective To investgate the effect of Tiao qi xing shui decoction combined with peritoneal injection DDP and IL-2 on Bax,Caspase-3, TGF-β1 gene expression in mouse with ascetes hepatoma,and explore its mechanism from molecular biology.Methods 72 pure lines of Kunming mice were randomly divided into control group,Chinese medicine group,combination of traditional Chinese and Western medicine,in high,middle,low dose group and Western medicine group,each had 12.All mices were killed after treatment,and the expression of Bax,Caspase-3,GF-β1 protein in mouse liver tumor tissues were detected by Immunohistochemical. Results Immunohistochemical results showed that,the positive expression of apoptosis gene Bax protein in high dose group of TCMand Western Medicine was higher than that of control group(P<0.01),and the positive expression of apoptosis gene Caspase-3,TGF-β1 protein in high and middle dose group of TCMand Western Medicine were higher than that of control group(P<0.01).Conclusion The high dose group of TCM and Western Medicine can significantly increase the expression of Bax protein,the high and middle dose guoup of TCMand Western Mdicine can significantly increase the expression of Caspase-3,TGF-β1 protein,induction apoptosis.

Objectives To explore the effects of hyperosmosis of sea water and low temperature on hemorrhage and coagulation sys- tem of dogs after open abdominal injury followed by sea water immersion,and to observe the efficiency of plasma in addition to routine first aid,so as to provide a theoretical basis for the early treatment of open abdominal wound in naval warfare.Methods 24 dogs with open in- jury of the abdomen were randomly divided to three groups with 8 animals for each group.The dogs in control group were not subjected to sea water immersion and received routine treatments;dogs in conventional therapy group were immersed in sea water and then given the routine first aid;dogs in plasma group were immersed with aqua marina and then given a combined treatment of routine first aid plus an in- fusion of plasma.The following parameters were measured and recorded:survival time,body temperature,mean arterial blood pressure, osmotic pressure of plasma,prothrombin time,partial thromboplastin time,d-dimer and factorⅡ.Results Prothrombin time and partial thromboplastin time were significantly prolonged,D-Dipolymer increased and factorⅡdecreased after sea immersion with open abdominal wound.The survival time of dogs in plasma group(45.6?12.5h)was significantly longer than that of dogs in both control and conven- tional therapy groups(P

Aim To investigate the pharmacokinetics of albumin-interferon ?-2b fusion protein(HSA-IFN?-2b) in Rhesus monkeys.Methods Three groups of monkeys were administrated by single injection of HSA-IFN?-2b at 50 ?g?kg-1 iv,50 ?g?kg-1 sc and 300 ?g?kg-1 sc,respectively.Blood samples were taken at 14 different time points from each animal.ELISA was employed to determine the drug concentrations in plasma for all samples.Results Drug was detectable in all of the treated monkeys up to 336 hours after administration.One-compartment model analysis showed that distribution volume was 71 ml?kg-1 in 50 ?g?kg-1 iv group,67 ml?kg-1 in 50 ?g?kg-1 sc group,and 63 ml?kg-1 in 300 ?g?kg-1sc group,respectively.The absorption of HSA-IFN?-2b was relatively slow when given by subcutaneous injection.After 50 ?g?kg-1 sc,the absorption half-life was 19h and elimination half-life was 53h,the bioavailability(F) was approximately 73%,and the apparent clearance(clearance divided by bioavailability,CL/F) was 0.92 ml?h-1?kg-1.Conclusion The result showed that HSA-IFN?-2b may offer the benefits of less frequent dosing and a potentially improved efficacy profile compared with non-albumin fused IFN-alpha.

Objective:To evaluate the efficacy and safety of pegylated liposomal doxorubicin (PLD) in CHOP regimen for un-treated elderly patients with advanced diffuse large B-cell lymphoma (DLBCL). Methods:In a prospective phase II study, we analyzed the feasibility of PLD-modified CHOP regimen in elderly patients with advanced stages of DLBCL. PLD was administered at 30 mg/m2 in combination with cyclophosphamide, vincristine, and prednisone at standard doses every 21 d for six cycles. CD20 positive patients were given option for rituximab treatment. Results:From November 2011 to March 2014, 30 patients with a median age of 70 years (range:63 to 80) were enrolled in this study. Up to 24 cases (80.0%) obtained an International Prognostic Index of≥3. The overall re-sponse rate was 86.7%, and the complete remission rate was 66.7%. With a median follow-up of 20.1 months, the 18-month overall and progression-free survival rates were 82.4%and 70.1%, respectively. The main toxicity was neutropenia, reaching grades 3 to 4 in the 24 cases (80.0%). No significant changes existed in patients' left ventricular ejection fraction and serum troponin-T during the study. Four patients (13.3%) showed asymptomatic abnormal changes in electrocardiogram after PLD infusion. Conclusion:CHOP regimen with PLD is an effective alternative for the treatment of DLBCL in elderly patients, exhibiting an acceptable toxicity.

Aim To evaluate the effects of ferulic acid ( FA ) on lipopolysaccharide ( LPS )-induced neuroin-flammation in microglia cells and its potential mecha-nisms. Methods Microglial activation was induced by stimulation with LPS, and the effects of FA pretreat-ment on microglial activation and production of proin-flammatory mediators, nitric oxide/iNOS were investi-gated. The role of the mitogen-activated protein kinases in the antiinflammatory actions of FA in LPS-stimulated microglia was further elucidated. Results Cell viabil-ity experiments revealed that FA did not produce cyto-toxicity in microglia. FA significantly inhibited LPS-in-duced production of tumour necrosis factor-alpha ( TNF-α) , interleukin-6 ( IL-6 ) , interleukin-1 beta ( IL-1β) , and nitric oxide ( NO ) . Protein and mRNA levels of COX-2 and inducible nitric oxide synthase ( iNOS) were also attenuated by FA. Further experi-ments on intracellular signalling mechanisms showed that inhibition of extracellular regulated kinase ( ERK) contributed to the anti-neuroinflammatory actions of FA. Conclusion The results suggest that FA inhibits LPS-induced microglial inflammation by partial targe-ting of ERK signalling and attenuation of ERK.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

OBJECTIVE: To learn the clinical features of patients infected with Klebsiella pneumoniae in our hospital.METHODS: From 2006 and 2007,a total of 96 inpatients in our hospital that were confirmed to be infected with Klebsiella pneumoniae by sputum culture were collected for a study of their clinical features and drug susceptibility test results.RESULTS: Patients with Klebsiella pneumoniae infection represented 12.7% of the total concurrent pulmonary infection cases,of which,57 Klebsiella pneumoniae infection cases had concurrent pulmonary tuberculosis(TB),and 39 Klebsiella pneumoniae infection cases had no concurrent TB.Drug susceptibility tests showed that the resistance rates of Klebsiella pneumoniae to third generation cephalosporins and quinolones were higher than those reported in the literature,and the resistant rate of Klebsiella pneumoniae to quinolones in Klebsiella pneumoniae infection patients complicated with TB was higher than those without TB.CONCLUSION: Klebsiella pneumoniae infection was more common in patients with low body resistance,which has typical clinical manifestations.Recently,the resistance of Klebsiella pneumoniae to the third generation cephalosporin has been increasing,and its resistance to quinolones in patients with pneumonia Klebsiella infection complicated with TB was significantly higher than in those without TB.

AIM: To investigate the effects of a 10-weeks treatment with angiotensin Ⅱ (Ang Ⅱ) subtype I receptor antagonist losartan on vascular remodeling of thoracic aorta in male spontaneously hypertensive rats (SHR). METHODS: SHR were treated from 16 to 26 weeks of age with losartan at 15 mg/kg?d -1 or 0.75 mg/kg?d -1. RESULTS: Losartan (15 mg/kg?d -1) treatment significantly decreased systolic blood pressure compared with the control group, while losartan (0.75 mg/kg?d -1) had no the effect, losartan(15 mg) prevents the development of aortic hypertrophy by preventing hypertrophy of vascular smooth muscle cells (VSMC). In the losartan 0.75 group, these parameters were not changed. But in the losartan 15 and losartan 0.75 groups, the collagen content of the aortic media decreased significantly. CONCLUSION: It is inferred that the effect of Ang Ⅱ on stimulating VSMC growth of the aorta in SHR is dependent on arterial pressure, while the effect on collagen fibers is through pressure independent mechanism.

AIM: To investigate the effects of angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension. METHODS: Eighteen 4 weeks old male rats were divided into three groups: Wistar-Kyoto rats (WKY) for normotensive group, and spontaneously hypertensive rats (SHR) for hypertensive group, and SHR treated with losartan orally (15 mg?kg -1 ?d -1 ). The rats were raised to 16 weeks old. The morphometric parameters of the renal arterioles, and the widths of vascular smooth muscle cells (VSMC) and intercellular space were studied on kidney slices by light microscope and electromicroscope respectively, combined with computer-assistant image analysis system. The minimal renal vascular resistance (RVR min ) was studied by isolated kidney perfusion system. RESULTS: The systolic blood pressure of the tail artery, wall thickness, wall area, ratio of wall thickness to inner diameter, width of VSMC of renal arterioles and RVR min were all smaller or lower in losartan group than those of SHR. CONCLUSION: Ang II receptor antagonist losartan can prevent the remodeling of renal arterioles in SHR.