Objective To determine the expression of hepatocyte growth factor and its receptor c-met in bladder transitional cell carcinoma, and to investigate correlations of the expression and clinic features of the tumor. Methods Samples of 45 human bladder carcinoma and 8 normal bladder tissues were detected with immunohistochemical method for HGF and c-met ,then relationships between the expressions and clinic features of the tumor were analyzed. Results There was no HGF expression and only 1 weak positive c-met expression in the normal urinary bladder tissues. While in bladde cancer specimens, positive expression rates of HGF and c-met were as high as 64.4%,62.2%, respectively.Coexpression rate of HGF and c-met was 42.2%. The positive rates were significantly higher in BTCC specimens than in normal tissues (P

Objective To construct tissue-engineered skin containing melanin with mixed culture of human keratinocytes (KCs) and melanocytes (MCs) on de-epidermized dermis (DED) in vitro. Methods Single-cell suspension was obtained by digestion of isolated preputial epidermis with pancreatin. Keratinocyte serum-free medium (K-SFM) and modified M254 culture medium were used to culture KCs and MCs respectively. Third-passage KCs were seeded into cell culture flasks and cultured for 48 hours; then, third-passage MCs were seeded into the same cell culture flasks with the MC/KC ratio being 1: 10 followed by a 5-day coculture. The suspension of third-passage KCs and MCs with the MC/KC ratio of 10:1 were seeded onto the surface of a prepared DED and maintained at the air-liquid interface for 11 days following a 4-day submerged culture.Subsequently, the constructed tissue-engineered skin was examined with HE staining, immunohistochemical staining for keratin and Masson-Fontana staining. Results After coculture in flasks for 5 days, KCs exhibited a typical paving-stone appearance, MCs with projected dendrites were scattered in the extracellular space between KCs. HE staining revealed 3 to 6 layers of cells with the formation of stratum corneum after mixed culture on DED for 15 days. Keratin protein was positive throughout the artificial epidermis, and melanin pigments were located in the basal layer of the epidermis as Masson-Fontana staining showed. Conclusions The co-culture of MCs and KCs can form single-cell layers with the contact between MCs and KCs in flasks, and construct tissue-engineered skin with melanin component on DED in vitro.

Objective: To extract the polysaccharides of Curcuma zedoaria Rosc (PCZ) and study its bioactivities. Methods: The crude polysaccharides were extracted from herbal slice of Curcuma zedoaria Rosc with hot water ,and precipitated by ethanol and savage deproteinization and depigmentation with activated charcoal. The constituents were isolated by column chromatography and the total polysaccharides content was determined. Experiments of immune function,anti-oxidation and anti-tumor activity of KM mice and in viro were studied. Results: The content of total polysaccharides of Curcuma zedoaria Rosc was 33.12%. and the extraction rate was 1.938% . It could raise spleen cell proliferation and transformation rate of lymphocyte and SOD activity in blood of mice. A higher antitumor effect of the polysaccharides by injection was observed dose-dependently. Conclusion: PCZ could raise the immune and anti-oxidative activities and then had a higher antitumor effect.

Objective To evaluate the effect of lovastatin on shedding of heparan sulfate proteoglycan (HSPG) and syndecan-1 (SDC-1) in the lung tissues of rats with sepsis-induced acute lung injury.Methods One-hundred and twenty male Wistar rats aged 8-12 weeks,weighing 325-425 g,were randomly divided into 3 groups (n =40 each) using a random number table:sham operation group (group S),cecal ligation and puncture (CLP) group and lovastatin group (group L).Lovastatin 4 mg/kg was injected once a day for 5 consecutive days in S and L groups,while the equal volume of 0.5％ CMC (the solvent) was given in CLP group.Sepsis was produced by CLP on 5th day of administration in CLP and L groups.The left lung was lavaged at 24 h after operation.The broncho-alveolar lavage fluid (BALF) was collected for determination of protein concentrations,white blood cell (WBC) count and percentage of neutrophils.Blood samples were collected for determination of the concentrations of HSPG and SDC-1 in serum (by ELISA).Evans blue was injected at 24 h after operation in the remaining 20 rats of each group.The lungs were removed for examination of the pathological changes and for measurement of HSPG and SDC-1 mRNA and protein expression (using Western blot and PCR),and Evans blue content (reflecting pulmonary capillary permeability) in the lung tissue.Results Compared with group S,the protein concentrations,WBC count and percentage of neutrophils in BALF,Evans blue content in lung tissues and the concentrations of HSPG and SDC-1 in serum were significantly increased,and HSPG and SDC-1 mRNA and protein expression was down-regulated in CLP and L groups.Compared with group CLP,the protein concentrations,WBC count and percentage of neutrophils in BALF,Evans blue content in lung tissues and the concentrations of HSPG and SDC-1 in serum were significantly decreased,and HSPG and SDC-1 mRNA and protein expression was up-regulated in group L.The pathological changes of lungs were significantly attenuated in group L as compared with group CLP.Conclusion The mechanism by which lovastatin attenuates acute lung injury induced by sepsis may be related to reduced shedding of HSPG and SDC-1 in lung tissues and improved function of pulmonary vascular endothelium in rats.

Aim Bupivacaine is a kind of long-acting amide local anesthetics.This paper aims to explore the effects of bupivacaine on the short-circuit currents in human alveolar epithelial monolayers and study the possible mechanisms.Methods Short-circuit currents were recorded by ussing-chamber setup.Amiloride-sensitive currents were defined as the difference be-tween the total current and the amiloride-resistant cur-rent.ERK1 /2 phosphorylation protein levels were ana-lyzed by Western blot at 0,1 5,30 and 60 min after administration of 1 00 μmol·L -1 bupivacaine.Results Bupivacaine could inhibit the short-circuit currents in H441 monolayers dose-dependently,which could be inhibited by amiloride.Western blot analysis showed that bupivacaine increased the level of ERK1 /2 phos-phorylation.Conclusion These data demonstrate that bupivacaine can reduce the alveolar ion transport by in-hibiting the amiloride-sensitive currents,possibly by the enhancement of ERK1 /2 phosphorylation. The effects of alveolar fluid clearance following application of bupivacaine should be considered clinically when the patient is complicated with lung injury.

Objective To compare effectiveness,performance,onset time and complications between ultrasound-guided cervical transverse process block and interscalene brachial plexus block in patients undergoing proximal humeral surgeries.Methods Sixty patients,27 males and 33 females, aged 18-70 years,scheduled for proximal humeral surgeries were randomly divided into two groups. They were given either cervical transverse process block (group T,n =30)or ultrasound-guided in-terscalene brachial plexus block (group I,n =30).All patients received a total of 8 ml of 0.5% ropiv-acaine.The performance, anesthetic onset time, the side effects and block success rate were evaluated.Results Block procedure was quicker in group T than in group I [(8.73 ±3.1 7)min vs. (14.40±8.21)min,P <0.01].The severity of diaphragmatic paralysis in group T was significantly lower than in group I (P <0.01).The ultrasound-guided cervical transverse process block was more effective than the interscalene brachial plexus block in patients undergoing proximal humeral surgeries (100% vs.80%,P <0.05).Conclusion The ultrasound-guided cervical transverse process block has a higher success rate and fewer incidence of diaphragmatic paralysis than the interscalene brachial plexus block in patients undergoing proximal humeral surgeries.

Objective To research the 50% effective anesthetic volume ( EAV50) of 0.5% ropivacaine in nerve stimulator-guided vertical infraclavicular brachial plexus block. Methods Thirty patients scheduled for forearm or hand surgery were blocked using 0. 5% ropivacaine in nerve stimulator-guided vertical infraclavicular brachial plexus block. The EAV50, which is the anesthetic volume corresponding to 50% success and 50% failure was determined by up-and-down sequential test. The starting dose of 0. 5% ropivacaine was 0. 55ml/kg. Block failure resulted in a dose increase 110% , and block success in a reduction 110%. The sensory and motor blockade were accessed at 5- min intervals (up to 60 min). Results In nerve stimulator-guided vertical infraclavicular brachial plexus block, EAV50 for 0.5% ropivacaine was 0.417ml/kg. In up-and-down sequential test, there were significantly different in the block success rates of lateral, medial and posterior cord ( P <0.01). Conclusion In nerve stimulator-guided vertical infraclavicular brachial plexus block, enough anesthetic volume must be given to increase block success rates of all the cords of brachial plexus, and EAV50 for 0. 5% ropivacaine was 0.417ml/kg.

Objective:To evaluate the value of adenosine triphophate (ATP)stress/rest nuclide myocardial perfusion imaging (MPI)in the diagnosis of female patients with coronary heart disease (CHD).Methods:The clinical materials of 47 female suspected CHD patients were retrospectively analyzed,aged from 39 to 74 years,and the average age was (53.7±6.3)years old.All patients were hospitalized and underwent two-day ATP stress and rest nuclide MPI and coronary angiography (CAG)in two weeks. The results and images of MPI and CAG were evaluated by more than 2 attending physicians. Using CAG as the “gold standard”, the diagnostic efficiency (sensitivity, specificity and accuracy) of MPI for CHD was evaluated. Results:Compared with CAG, the sensitivity,specificity and accuracy of ATP stress MPI in diagnosing the female CHD patients were 81.3% (13/16),77.4% (24/31)and 78.7% (37/47)individually;the positive predictive value and negative predictive value were 65.0% (13/20)and 88.9% (24/27).There were no severe adverse effects in the ATP stress test and the incidence of adverse effects was 85.1%.Conclusion:There is a highly diagnostic efficiency of ATP stress MPI in the CHD patients.It can be the first choice of examination methods for screening without injury and diagnosing the myocardial ischemia in the female patients.

Objective:To evaluate the role of ten-eleven translocation methylcytosine dioxygenase 3 (TET3) in trigeminal ganglion in maxillofacial inflammatory pain in mice.Methods:Forty SPF healthy male C57BL/6J mice, aged 8-10 weeks, weighing 19-23 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), inflammatory pain group (group IP), control+ TET3-siRNA group (group C+ siTET3), inflammatory pain+ TET3-siRNA group (group IP+ siTET3) and inflammatory pain+ negative control Scrambled-siRNA group (group IP+ siNC). Normal saline or complete Freund′s adjuvant (CFA) 10 μl was injected into the temporomandibular joint of mice, respectively, and the mechanical paw withdrawal threshold (MWT) was measured at 1, 4, 8 and 12 days after injection (T 1-4). Before injection of normal saline or CFA, 0.75 μl siTET3 or siNC was injected into the trigeminal ganglion and the animals were then sacrificed and trigeminal ganglion was removed at T 2 for determination of the expression of TET3 by Western blot in C+ siTET3, IP+ siTET3 and IP+ siNC groups. Results:Compared with group C, MWT was significantly decreased at T 1-3 , the expression of TET3 in trigeminal ganglion was up-regulate in group IP ( P<0.05 or 0.01). Compared with IP and IP+ siNC groups, MWT was significantly increased at T 2, 3, and the expression of TET3 in trigeminal ganglion was down-regulate in group IP+ siTET3 ( P<0.05 or 0.01). Conclusion:TET3 in trigeminal ganglion is involved in the development of maxillofacial inflammatory pain in mice.

AIM: Through studying the differences between wild-type acidic fibroblast growth factor (waFGF) with recombinant aFGF (raFGF), the biological effect of raFGF concerned with mitogenic activity was evaluated. METHODS: NIH3T3 cell line was used. Cell proliferation with MTT method was used to study the mitogenic activity. Flow cytometry was also used for detection of apoptosis, cell membrane permeability and mitochondria potential. The role of heparin sulfate (HS) on aFGF biological effect was studied at the same time in this research. RESULTS: The enhancement of raFGF on cell proliferation was significantly lower than that of waFGF. The restriction of raFGF on apoptosis and the enhancement of it on cell membrane permeability were all lower than those of waFGF significantly. The enhancement of raFGF on mitochondria potential was lower than that of waFGF significantly. The HS improved the biological effect of aFGF. CONCLUSION: The mitogenic activity of raFGF is lower than that of waFGF and raFGF has little effect on apoptosis.