Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.
Apoptosis ; Cell Proliferation ; Dependovirus ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; Serpins ; metabolism ; Signal Transduction ; Transfection

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

Danshen,an efficacious agent for cardiovascular diseases,has been found to play an essential role in kidney injury.In the present study,the effect of Danshen on cisplatin-induced renal dysfunction was investigated in a mouse model.Danshen was administered to mice at a dose of 3 g/kg 4 days before and 3 days after cisplatin treatment.A single intraperitoneal injection of 20 mg/kg cisplatin was used to induce nephrotoxicity.The mice were sacrificed 72 h after cisplatin intoxication.Biochemical parameters including serum creatinine and blood urea nitrogen were analyzed.Histopathological changes of kidney tissues were detected using HE staining.Antioxidant enzymes (GSH-Px and SOD) and peroxidative product (MDA) were detected.Protein expressions of Nrf2 and its target genes including HO-1 and NQO1 were measured by Western blotting.The results showed that pretreatment with Danshen significantly reduced serum creatinine and blood urea nitrogen in the cisplatin-treated mice.Histopathological examination showed that Danshen mitigated the renal damage induced by cisplatin.Moreover,Danshen restored the activities of antioxidant enzymes (GSH-Px and SOD) and normalized the MDA contents in renal tissues.Western blotting revealed that Danshen enhanced the expressions of Nrf2 and its target genes in cisplatin-exposed mice.It was suggested that Danshen protects against the cisplatin-induced renal impairment in the mice,which is potentially associated with the upregulation of Nrf2-mediated signaling pathway.

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

Objective To investigate HPV infection and its correlation with the expression of sur- vivin and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA).Methods HPV6/11 and 16/18 DNA were detected by PCR in 41 cases of CA and 23 specimens of normal skin.The expression of survivin and PCNA was measured by immunohistochemical technique in these two groups. Results The infection rates of HPV6/11 and 16/18 were 87.80% (36/41) and 19.51%(8/41),respective- ly,in CA,and 0 in normal skin epidermis.The expression of survivin and PCNA was found in 53.66% (22/41) and 87.80% (36/41) of CA ,and was 8.70%(2/23)and 47.83% (11/23) in normal skin,respec- tively.The expression of survivin was positively correlated with the high expression of PCNA in CA.The expression of survivin and PCNA was higher in HPV6/11 positive lesions than in HPV6/11 negative lesions. Conclusion HPV infection may up-regulate the expressions of survivin and PCNA,inhibit the apoptosis and promote the proliferation of keratinocytes.

OBJECTIVETo review the long-term clinical effect of composite transplantation of allogeneic acellular dermal matrix (ADM) and split thickness skin autograft (STSG).

METHODSNineteen patients with 34 wounds transplanted with allogeneic ADM combined with STSG who were hospitalized from March 2001 to October 2008 were enrolled as composite transplantation group (CT). Another 9 patients with 11 wounds transplanted with STSG admitted within the same time frame were enrolled as control group (C). All patients were followed up for longer than 2 years. Color, evenness, texture, contracture, sensation, and complications of transplanted skin were assessed using a modified Manchester Scar Scale (1-4 scores, the higher the score, the poorer the situation). The scar formation on skin donor sites was assessed by the Vancouver Scar Scale. Patients' degree of satisfaction and health status during the transplantation period were investigated in the form of questionnaire. The skin tissue structure of 4 patients was observed with histological method. The joint range of motion was assessed by the neutral position before and after operation and at follow-up. Data were processed with nonparametric test, chi-square test or t test.

RESULTS(1) The evenness, contracture, and texture of transplanted skin in CT group scored (1.6 ± 0.5), (1.8 ± 0.8), and (1.5 ± 0.8), respectively, which were significantly lower than those in C group [(2.0 ± 0.7), (2.2 ± 0.9), and (2.3 ± 0.7), with Z value respectively -2.058, -2.220, -2.323, P values all below 0.05]. Scores of color, sensation, and complications of transplanted skin in two groups were close to each other (with Z value respectively -0.628, -0.428, -2.520, P values all above 0.05). (2) Mild scar formation was observed in one of the skin donor sites in CT group. (3) Information as obtained from questionnaire showed no statistical difference between two groups in pinching, itching, and satisfaction degree (with χ(2) value respectively 0.187, 0.019, 2.628, P values all above 0.05). (4) Nerve fibers were seen in hand tissue 2 years after operation. ADM did not induce severe inflammatory responses in the site of grafting. (5) Eleven joints in CT group recovered or improved in function; while the other two joints required secondary surgery. Obvious contracture was observed in the two joints in C group.

CONCLUSIONSAllogeneic ADM combined with STSG transplantation prevents scar contracture and has obvious effect in improving function and appearance. There is no problem in regard to safety for its existence in either adult or children.

Adolescent ; Adult ; Burns ; surgery ; Child ; Child, Preschool ; Dermis ; transplantation ; Female ; Follow-Up Studies ; Humans ; Male ; Skin Transplantation ; methods ; Skin, Artificial ; Time ; Transplantation, Autologous ; Transplantation, Homologous ; Young Adult

AIMTo investigate whether NR2B-pERK1/2-pElk-1 signaling contributes to the Y-maze learning and memory of rat brain.

METHODS45 adult male SD rats were divided into 4 groups: (1) Ifenprodil peritoneal injection group (Ifenprodil ip, n = 14); (2) DMSO peritoneal injection group(DMSO ip, n = 15); (3) Ifenprodil cerebral ventricle injection group (Ifenprodil ic, n = 8); (4) DMSO cerebral ventricle injection group(DMSO ic, n = 8). Y-maze training and test were used as an learning and memory enhancing stimulus. Immunohistochemical and Western blotting methods were used for detecting pERK1/2 and pElk-1 expression intensity of different brain regions.

RESULTSCompared with the DMSO ip group, the ifenprodil ip group showed no change on the Y-maze learning score (P > 0.05), but its Y-maze memory score tested 24 after learning decreased (P < 0.05). Ifenprodil peritoneal injection made brain pERK1/2 and pElk-1 expression decreased generally. In hippocampus, marginal division of striatum(MrD), amygdala,these changes were more significant (P < 0.05). Compared with the DMSO ic group, the reconsolidation of Y-maze memory tested 6 hours after ifenprodil injection was impaired in ifenprodil ic group (P < 0.05). The OD value of pERK1/2 and pElk-1 positive bands in ifenprodil ic group attenuated generally. The pElk-1 positive bands of caudate putamen and MrD almost disappeared in ifenprodil ic group.

CONCLUSIONNR2B is essential for the formation of long-term memory, reconsolidation of Y-maze memory. The deactivation of NR2B by ifenprodil will impair these courses. Meanwhile, the deactivation of NR2B attenuates pERK1/2 and pElk-1 expression of learning and memory related regions after Y-maze learning and memory reconsolidation test. In MrD and caudate putamen, the pElk-1 expression are completely blocked by ifenprodil after memory reconsolidation test.

Animals ; Avoidance Learning ; physiology ; Dimethyl Sulfoxide ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Piperidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; ets-Domain Protein Elk-1 ; metabolism

OBJECTIVETo investigate the bacterial flora and their drug resistance in hospitalized burn patients on tropical islands.

METHODSRetrospective study was carried out to analyze pathogenic microorganisms and their drug resistance characteristics in 392 burn patients hospitalized during 2000-2005.

RESULTS(1) Totally 671 strains of bacteria were isolated, among which Pseudomonas aeruginosa, Staphylococcus aureus, Aerobacter cloacae and Acinetobacter species were predominant, but the isolation rate of Pseudomonas aeruginosa was declining compared with that in 1990's. (2) The resistance rate of Pseudomonas aeruginosa to imipenem and cefepime was 32.2% and 36.7% , respectively, while that to other antibiotics was above 80%. (3) Among 141 strains of Staphylococcus aureus, 89 strains (63.1%) of MRSA were isolated, and none of them were resistant to Vancomycin.

CONCLUSIONPseudomonas aeruginosa, Staphylococcus aureus, Aerobacter cloacae and Acinetobacter species were predominant in tropical islands. The species of the pathogens are changing . The drug resistance rates of Pseudomonas aeruginosa and Staphylococcus aureus are rather high.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; pharmacology ; Burns ; drug therapy ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Retrospective Studies ; Staphylococcus aureus ; drug effects ; isolation & purification ; Tropical Climate ; Young Adult

OBJECTIVETo sum up the clinical experience in the reparation of burn wounds with different types of skin flaps in patients with severe electrical injury.

METHODSFree skin flap, axile island flap with vascular pedicle and muscular skin flap were employed to repair 64 wounds in 49 cases.

RESULTSSkin flap necrosis happened in 2 cases, congestion and necrosis at the edge of skin flaps in 4 cases, and complete survival of the skin flaps and primary healing of the wounds in the remaining cases.

CONCLUSIONReparation of wounds with proper skin flaps according to the degree of the electrical injury is effective and reliable, which might be beneficial to the prevention of necrobiosis and the infection in deep tissue.

Adolescent ; Adult ; Burns, Electric ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps

OBJECTIVETo observe the changes of auditory steady-state response (ASSR) threshold in guinea pigs with posterior semicircular canal fistula.

METHODSIn 10 guinea pigs, a window was opened in the posterior semicircular canal of the left ear with the right ear serving as the control for testing the ASSR under anesthesia with pentobarbital sodium.

RESULTSThe mean and standard deviation of ASSR threshold (dB SPL) at 0.5, 1, 2, and 4 kHz in the left ear was 35.00+/-14.33, 25.50+/-12.37, 20.00+/-9.37 and 20.00+/-9.18, respectively, and was 31.00+/-16.19, 25.50+/-12.34, 18.00+/-6.96 and 18.50+/-6.71 in the right ear, respectively. Paired-sample t test showed no significant difference in the ASSR at the same frequency between the two ears.

CONCLUSIONSmall fistula by surgery causes no significant hearing loss in guinea pigs.

Animals ; Audiometry, Pure-Tone ; methods ; Auditory Threshold ; physiology ; Cochlea ; pathology ; Evoked Potentials, Auditory ; Fistula ; pathology ; physiopathology ; Guinea Pigs ; Labyrinth Diseases ; pathology ; physiopathology ; Semicircular Canals