AIM: To study the effect of Lomerizine on the activity of P-glycoprotein (P-gp) in the bloodbrain barrier(BBB) and search for novel effective P-gp inhibiting agent against multidrug resistance. METH-ODS: Rhodamine123 (Rh123) was used to examine the activity of P-gp and RT-PCR to study the mdr mRNA expression in cultured rat brain microvessel endothelial cells (RBMECs). RESULTS: Lomerizine could increase the cellular Rh123 in RBMECs in a concentration-dependent manner. RT-PCR indicated that lomerizine could not down-regulate the expression of mdr mRNA. CONCLU-SION: Lomerizine can reverse multidrug resistance in the blood-brain barrier by inhibiting the activity of P-gp.KEY WORDS lomerizine; P-glycoprotein; bloodbrain barrier; RT-PCR

AIM: To observe protective effects of sanguis draxonis flavones on myocardial ischemia in rats and dogs. METHODS: Acute myocardial ischemia in rats was produced by iv pituitrin and ECG indexes were observed. Myocardial ischemia was induced by ligating coronary artery in anaesthetized dogs, and then EEC, CK,LDH, and LD in serum were determined respectively. RESULTS: J point and T wave in rats changed evidently after iv pituitrin, which was reversed by sanguis draxonis flavones (360, 180 mg?kg~ -1). In coronary artery ligation model, infraction range, △N-ST, △?-ST and some serum indexes (such as CK, LDH and LD) was decreased after ig sanguis draxonis flavones (120, 60, 30 mg?kg~ -1). CONCLUSION: Acute myocardial ischemia is protected effectively by sanguis draxonis flavones.

Immunologic classification of acute lymphocytic leukemia (ALL) was carried out by using 11 - 13 kinds of monoclonal antibodies against human leucocyte differentiation antigens in 19 cases. The results showed that of these 19 cases, there were T-ALL in 4, B-ALL in 4, C-ALL in 7, AUL in 2 and hybrid type of ALL (H-ALL) in 2. The 5 types mentioned above could also be classified, according to the immunologic criteria described by Foon in 1986, into 3 major groups: T-ALL, non-T-ALL and H-ALL. The Foon's classification method is useful for judgement of malignant cell source and differentiated stage, and for further understanding the nature of ALL cells.

Objective To evaluate the effects of external substance P (SP) on scalding wound healing and neovascularization.Methods Eighty-four Wistar rats were induced into diabetic models by intraperitoneal injection of streptozotocin (STZ), and deep partial thickness scalding wound on the back with diameter of 2 cm was prepared. Rats were randomly divided into experiment group (n=42, local injection of SP) and control group (n=42, local injection of PBS). The process of wound healing was observed, and the percentages of wound closure were calculated on D0, D1, D3, D7, D10, D14, D21 post scald. The expression of CD31, surface marker of neovascular endothelial cells, was detected within the wound sites by immunohistochemical staining, and the microvessel density was calculated. Results The percentage of wound closure in experiment group was significantly higher than that in control group on D7 post scald [(42.69±3.26) % vs (30.24±1.17)%, P<0.01]. Immunohistochemical detection revealed that the expression of CD31 and the microvessel density in experiment group were significantly higher than those in control group from D7 post scald (P<0.01). ConclusionExternal SP may promote scalding wound healing in diabetic rats, the mechanism of which may be associated with upregulation of expression of CD31 and acceleration of neovascularization.

This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.

Objective To evaluate the clinical efficacy of acupoint injection with BCG-polysaccharide nucleic acid (BCG-PSN) in treating bronchial asthma. Method Seventy-two patients with bronchial asthma were randomized into an acupoint injection group and a muscular injection group, 36 cases in each group. The acupoint injection group was intervened by acupoint injection with BCG-PSN to Zusanli (ST36) and Dingchuan (EX-B1) alternately, 1 mL for two points in total each time; the muscular injection group was intervened by muscular injection at the same dose and frequency, twice a week, for successive 3 months. The pulmonary function and asthma control test (ACT) were estimated before and after intervention and during March of the next year. Result After intervention, the FEV1 values were (80.97±2.31)% and (80.78±2.56)% respectively in the acupoint injection group and muscular injection group, and PEF values were (6.50±0.21)L/s and (6.48±0.25)L/s, and the between-group differences were statistically insignificant (P>0.05). The ACT score was (23.02±1.03) in the acupoint injection group, significantly better than (22.40±2.04) in the muscular injection group (P<0.05). The follow-up study showed that the ACT score in the acupoint injection group was superior to that in the muscular injection group (P<0.05). Conclusion Acupoint injection and muscular injection with BCG-PSN can equally improve the pulmonary function in bronchial asthma, while the acupoint injection can produce a more significant effect than muscular injection in improving ACT.

Aim: To evaluate the effects of HZ08, a novel P-glycoprotein inhibitor, on reversing tumor resistance of K562/ADM to adriamycin in nude mice and on the activities of cytochromes P-450 (GYP) isoforms. Methods: Nude mice bearing K562/ADM were injected at different doses of HZ08 with adriamycin for 4 weeks. The tumor weights of HZ08 treatment groups were determined and compared to those of the control and positive groups. In addition, the effects of HZ08 were examined on GYP isoforms-mediated metabolism of specific substrates by GYP isoforms in rat liver microsomes in the presence or absence of HZ08. Results: The tumor weights of HZ08 treatment groups were significantly decreased and HZ08 was a relatively potent inhibitor of CYP3A4, with no significant effects on other isoforms tested. Conclusion: HZ08 has potent effects on reversing P-glycoprotein mediated tumor multidrug resistance in rive with little influence on cytoehrome P-450 activities of rat liver.

ObjectiveTo develop a new sutureless technique (photochemical tissue bonding,PTB ) for repair of corneal perforation. Methods A total of 60 rabbits were used for creating corneal perforation models.The corneal perforation on the left eye was repaired by sutures and the injury on the right eye was fixed with the use of amniotic membrane with PTB.The outcomes of the two mentioned repair methods were compared by observing the leakage of aqueous and the morphology of the anterior chamber at different instants,measuring the intraocular pressure (IOP) and observing the formation of neo-vessels and scars of cornea in the use of histological analysis. Results There was no leakage of aqueous and no difference for morphology evaluation in both treatments.PTB could adhere AM on the cornea to restore the corneal perforation.The peak IOP in the PTB treatment group at days 0,7 and 14 postoperative [ (531.2 ±49.5) mm Hg,(542.6 ±74.8) mm Hg and (603.9 ±69.1) mm Hg,respectively] was significantly higher than that in the suture group at the same instants [ (41.3 ±12.7) mm Hg,(142.6 ±25.4) mm Hg and (333.3 ± 66.7) mm Hg,respectively] (P ＜0.O1 ).Compared with suture repair,the treatment with PTB resulted in a better outcome of wound healing with less neo-vessels and less scars of cornea. Conclusion PTB treatment for repair of corneal perforation is superior to suture repair.

Objective To study the effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on the expression of vascular endotllelial growth factor (VEGF) in human dermal fibroblast. Methods In vitro human dermal fibroblasts in good status were incubated with GM-CSF (GM-CSF group) or non-GM-CSF (control group) culture medium for different periods of time. The mRNA, protein expression of VEGF in derma fibroblast were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively, and the secretion of VEGF in supernatant was measured by enzyme linked immunosorbant assay (ELISA). Results The expression of VEGF mRNA from dermal fibroblasts was increased significantly after l or more hours of incubation with GM-CSF comparing with the control (P＜0.05). 6 hours of stimulation by GM-CSF caused maximal expression of VEGF mRNA. The expression of VEGF protein in dermal fibroblasts was increased from 12 hours and was peaked at 24 hour after stimulation by GM-CSF. VEGF protein from the supernatant of the dermal fibroblasts was also raised persistently from 12 hour after stimulation by GM-CSF and was improved remarkably compared with the control. Conclusions GM-CSF can up-regulate directly the expression of VEGF in human derma fibroblast, which may be one of the mechanisms that GM-CSF accelerates neovascularization in wound healing.

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.