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 evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P＜0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P＜0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P＞0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P＞0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P＜0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P＜0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P＞0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.

Objective To observe ultrastructure of the morphological relationship between neuron and astrocytes in hippocampi of pentylenetetrazol (PTZ)-kindled epileptic rats,and to investigate the communicative ways between them.Methods Epilepsy models of 10 kindled rats established by intraperitoneal injection of PTZ[i.p.,35 mg/(kg · d)],assigned as kindled group.Five rats received 9 g/L saline as control group.Three days after being kindled,the ultrastructural relationship between neurons and the astrocytes was observed with transmission electron microscope and Cx43 labelling immuno-electron microscopy.Results 1.Synapses increased in hippocampi of PTZ-kindled epileptic rats.2.Gap junctions were observed between astrocytes and neurons.3.Astrocytic process extended into the synaptic cleft between pre-synaptic and post-synaptic membranes which formed the synaptic complex.4.The Cx43-hemichannels existed between astrocytes and neurons.Conclusions In hippocampi of PTZ-kindled epileptic rats,ultrastructure of morphological relationship between neurons and astrocytes includes synaptic complex,gap junctions and hemichannels,which might be communicative forms between neurons and astrocytes.

Group A rotavirus (RV) is the most important etiologic agent of severe gastroenteritis among children and the development of an effective vaccine becomes the top public health priority. Since survey of RV serotypes circulating in local community is important for introduction or development of RV vaccine, RV serotype G3 had proved as the predominant strain in Changchun from 2001 to 2005. Stool specimens collected from children with acute diarrhea were tested for group A rotavirus by enzyme-linked immunosorbent assay (ELISA) and RV isolates were typed by reverse transcription-polymerase chain reaction (RT-PCR) using serotype-specific primers. The complete VP7 gene segments of 31 rotavirus strains selected in Changchun from 1999 to 2005 were amplified with RT-PCR. Amplicons were cloned and sequenced. Comparative analysis of the VP7 sequences showed that there were no obvious differences among 31 RV strains. There was similar genetic variation among VP7 genes during the same RV season. The nucleotide sequence of VP7 gene of six G3 RV strains had one base deletion at nt1038 in 2003 RV season. The nucleotide mutations in regions A, B and C of VP7 gene took place at the same position or position near-by. Increase of nucleotide mutation in non- high variation region may benefit maintenance of serotype G3 as pre dominant strain after 2002. Increase of non continuous variation in non-high variation regions was notable.
Antigens, Viral ; genetics ; Capsid Proteins ; genetics ; Genetic Variation ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Serotyping

OBJECTIVETo investigate the risk factors related to outcome of chronic severe hepatitis B.

METHODSA total of 336 consecutive patients with chronic severe hepatitis B (CSHB) were analysed retrospectively. According to the outcome, objects were divided into survival group (n = 137) and death group(n = 199), then to observe the differences between them in respect to age, sex, family history, prothrombin activity (PTA), complications including ascites, infection, electrolyte disturbance, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal syndrome and the corresponding quantity of complications in each individual, antivirus therapy, artificial liver support system (ALSS) therapy, and alprostadil therapy. Finally, risk factors related to prognosis were selected by stepwise Logistic regression analyse.

RESULTSIn univariate analyse, significant differences between the two groups were found related to age, PTA, complications and its quantity (P < 0.01 for all), and antivirus therapy (P < 0.05) rather than sex, family history and treatment of ALSS or alprostadil. Logistic regression revealed that risk factors comprised of PTA and quantity of complications, antivirus therapy was the only protective factor.

CONCLUSIONA numbers of factors including age, PTA, complications and its quantity, and antivirus therapy affect the prognosis of CSHB, among which, antivirus therapy can reduce the death rate.

Adult ; Female ; Hepatitis B, Chronic ; diagnosis ; Humans ; Logistic Models ; Male ; Prognosis ; Retrospective Studies ; Risk Factors ; Treatment Outcome

Traumatic brain injury (TBI) is a highly complex multi-factorial disorder. Animal models of TBI are used to elucidate primary and secondary injury mechanisms and pathophysiological changes and to provide the diagnostic and therapeutical basis for TBI. The choices of animal models depend upon the research objectives. However, various animal models have limitations. The models only can duplicate the pivotal injury mechanisms or a certain important pathophysiological course. The characteristics of human TBI can not fully be reflected by using these models. In the review, animal models of traumatic brain injury are classified as dynamic direct brain injury, indirect dynamic brain injury and combined neuro-traumatic models. Several common models are described for consideration.
Animals ; Biomechanical Phenomena ; Brain/physiopathology* ; Brain Injuries/physiopathology* ; Diffuse Axonal Injury/physiopathology* ; Disease Models, Animal ; Forensic Medicine ; Head Injuries, Closed/physiopathology* ; Head Injuries, Penetrating/physiopathology* ; Humans ; Mice ; Rats ; Reproducibility of Results

To develop and optimize a simultaneous detection method of RotavirusA, Norovirus GI, GII, Sapovirus, human astrovirus, enteric adenoviruses and HBoV2 with GenomeLab GeXP analysis system. The sensitivity was verified to be 10(4) copies/microL with plasmids containing the viral targets in triplicate on different days, and no cross-reaction with enterovirus71, human Parechovirus and PicobirnavirusII was observed. Finally, we successfully developed a high throughout, rapid and maneuverable multiplex RT-PCR assay for simultaneous detection of seven viruses related with viral gastroenteritis, which provide a novel method for the molecular diagnosis of diarrhea-associated virus.
Gastroenteritis ; virology ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Viruses ; isolation & purification

OBJECTIVETo investigate whether the p38 mitogen-activated protein kinase (MAPK) signaling pathway mediates advanced oxidation protein products (AOPPs)-induced epithelial-to-mesenchymal transition (EMT) in tubular cells.

METHODSHuman proximal tubular cells (HK-2 cells) exposed to AOPP-bovine serum albumin (BSA) were examined for expressions of p38 MAPK and phosphorylated p38 MAPK using Western blotting. Western blotting and quantitative RT-PCR were used to examine the protein and mRNA expressions of EMT markers E-cadherin and vimentin and endoplasmic reticulum stress marker glucose-regulated protein (GRP) 78 in cells treated with SB203580 (an inhibitor of the p38 MAPK signaling pathway) prior to AOPP exposure. The cells treated with AOPPs following pretreatment with salubrinal (an inhibitor of endoplasmic reticulum stress) were also examined for expressions of p38 MAPK and phosphorylated p38 MAPK.

RESULTSAOPP treatment induced the phosphorylation of p38 MAPK in HK-2 cells. AOPP-induced decrease in E-cadherin expression and overexpression of vimentin and GRP78 were partly inhibited by pretreatment of the cells with SB203580. Salubrina partly suppressed AOPP-induced phosphorylation of p38 MAPK in the cells.

CONCLUSIONp38 MAPK signaling pathway, which is regulated by endoplasmic reticulum stress, might mediate AOPP-induced EMT in HK-2 cells.

This study was designed to evaluate the anti-inflammatory effect of recombinant human kallistatin (Kal) on ulcerative colitis (UC) in the mouse model. Acute colitis was induced by administration of 4% dextran sodium suffate (DSS) to KM mice for 7 days. The mice were then randomized into 5 groups:model control, Kal 0.2 mg·kg^-1·d^-1, 1.0 mg·kg^-1·d^-1 and 2.0 mg·kg^-1·d^-1 group, salazosulfapyridine (SASP) group. Ten age-matched normal KM mouse were administered with saline in the normal control. The weight, colon length, inflammation factor (MPO/SOD/MDA) and TNF-α/IL-10 levels among the five groups of mice were determined. The results showed that histological index score and MPO/MDA/TNF-α levels of high-dose Kal treatment group and SASP group were significantly lower compared with the model group (P<0.01), but the weight, colon length, IL-10 level and SOD activity were significant higher than the model group (P<0.01), approaching the normal group. These parameters showed that Kal can significantly relieve the UC state in a dose-dependent manner. This study demonstrates that Kal significantly remits UC in mice, and participates in the regulation of inflammatory cytokines TNF-α/IL-10 levels and has some antioxidant activity.

The present study was aimed to investigate the underlying mechanisms of the protective effect of proanthocyanidin (Pro) against hypoxia/reoxygenation (H/R) injury in H9C2 cells with a focus on Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway. H9C2 cells were randomly assigned to 5 groups, including the control group (Con), the H/R-injured group (H/R), the Pro-treated group (H/R+Pro), the JAK2 siRNA-treated group (H/R+Pro+JAK2 siRNA) and the JAK2 siRNA control group (H/R+JAK2 siRNA). The cells were pretreated with Pro (40 µmol/L) for 8 h before 2 h of hypoxia and 4 h of reoxygenation. Cellular viability and apoptosis rate were detected by MTT and TUNEL methods, and superoxide generation was measured. JAK2/STAT3 signaling, oxidative stress markers and endoplasmic reticulum stress markers were also detected by Western blot. We found that Pro treatment significantly improved cellular viability and reduced apoptosis rate in H/R-treated H9C2 cells. In addition, Pro treatment significantly up-regulated the phosphorylation levels of JAK2 and STAT3, down-regulated the superoxide generation, gp91, glucose-regulated protein 78 (GRP78), CCAAT/enhancer binding protein homologous protein (CHOP) and caspase-12 expression. However, these protective effects of Pro were all attenuated by JAK2 siRNA administration. Taken together, we demonstrated that Pro protects H9C2 cells against H/R-induced oxidative stress and endoplasmic reticulum stress injury via JAK2/STAT3 signaling pathway.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Cell Survival ; Endoplasmic Reticulum Stress ; In Situ Nick-End Labeling ; Janus Kinase 3 ; Oxidation-Reduction ; Phosphorylation ; Proanthocyanidins ; Protective Agents ; RNA, Small Interfering ; Rats ; STAT3 Transcription Factor ; Signal Transduction ; Up-Regulation