Aim To evaluate the effects of fibrinolytic enzyme FⅡ from agkistrodon acutus venom on an experimental model of kidney thrombus induced by lipopolysaccharide(LPS). Methods The model of microvascular thrombosis in the rabbits kidney was performed by the method of Hermida, which was induced by infusing LPS. Treatments were begun simultaneously with LPS infusion, through the contralateral marginal ear vein. Six different groups were established: NS 10 ml?h~-1 was infused as the negative control group, urokinase ~20 000 IU?kg~-1 ?h~-1 as positive control group, FⅡwas infused with the dosage of 0.1(Low-dose), 0.3 (medium-dose),0.6 (high-dose) mg?kg~-1 ?h~-1 . The further rabbits, which were given neither LPS nor FⅡ, were infused with saline solution through both marginal ear veins. Kinney sections were examined for the presence of fibrin microthrombi. The measurement of FDP concentrations was used to assess the degradation of microvascular thrombosis. Results Intense fibrin deposition was also detected and FDP concentrations were (78.21?4.79)% and (84.27?6.21)% at 2 and 6 hours after LPS administration in LPS-control group. Little fibrin deposition was detected and FDP concentration also increased in urokinase control group. A lot of fibrin deposition was detected in Low-dose FⅡ group,little fibrin deposition was detected in medium-dose FⅡ group, and no fibrin deposition was detected in high-dose FⅡ group. Additional all doses of FⅡ led to a significant increase in FDP concentration as compared with LPS-control group (P

AIM: To investigate the inhibitory effect of Chinese medicine Jinan injection(JA) on Lewis lung cancer (LLC) in mice. METHODS: The C 57 BL/6J mice with Lewis lung cancer(LLC) were divided into normal saline(NS), Jinan high dose (JAH), Jinan middle dose (JAM), Jinan low dose (JAL) and cyclophosphamide(CTX) groups. The body weight changes and inhibitory rate of LLC in each group were observed. In addition, flow cytometry and TUNEL were used to detect the anticancer mechanism of Jinan. RESULTS: The body weights were increased significantly in JA-treated groups vs CTX and the resistant rate was 45.79%, 40.90%, 32.48% and 98.96%, respectively. The apoptotic rate was 24.19%, 14.95% and 13.93% in JAH, JAM and JAL, respectively, and the Jinan induced apoptosis of LIC in a dose-dependent manner.CONCLUSION: Jinan injection inhibites the growth of LLC, and the apoptosis induction may be one of mechanisms that Jinan treates LLC in mice.

[Objective] To purify and characterize a novel factor X activator,Fve-1 from Daboia russelli siamensis (Myanmar) venom.[ Methods]F V e-1 was purified by ion-exchange chromatography and gel filtration.The hemostatic activity of F V e-1 was determined based on chromogenic substrates.The fibrinogen-clotting activity of F V e-1 was also determined.Thermal stability, pH stability,enzyme activity,and inhibition of F V e- 1 were determined by its remaining procoagulant activity.N-treminal sequence was determined by the method of automated Edman degradation.[ Results ]F V e-1 was achieved by chromatography with a molecular weight of 13,808 and an isoelectric point of 4.6. The hemostatic activity of 0.5 mg Fve-1 was equal to that of 1.5625 u thrombin or that of 54.93 ng RVV X. F V e-1 primarily activated F X, but did not affect on prothrombin and fibrinogen. The suitable pH and temperature range of F V e-1 was 6.5-7.5 and 25-60 ℃,respectively.The activity of F V e-1 was enhanced by Ca2+ and inhibited by EDTA and DTT.The N-terminal sequence of F V e-1 was NH2-N-L-Y-Q-F-G-E-M-I-N.[Conclusion] F V e-1 is a factor X-activating enzyme,which could activate FX to FX a,but have minimal effect on prothrombin and fibrinogen.

Objective To evaluate the clinical effect of computed tomography?guided visceral nerve plexus ethanol neurolysis through post?curs of diaphragm approach in the treatment of patients with pancreatic cancer pain using,and study the safety and life quality improvement of patients . Methods A total of 58 patients suffered from pancreatic cancer pain,who were treated in the department of pain medicine of the First Hospital of China Medical University from October 2013 to December 2014,were recruited for the study. The patients were divided into two groups according to the willing of the patients and their families,group A(32 cases)was treated with Visceral Nerve Plexus ethanol Neurolysis,while group B(26 cas?es)was treated with oral opioid drugs. The analgesic effect,changes in the amount of opioid drugs,changes in the PSQI scores and the improvement of quality of life were evaluated before treatment and 1 day(T1),15 days(T15),30 days(T30),60 days(T60)after treatment. Record the adverse reactions in the course of treatment. Results All the patients of group A successfully received visceral nerve plexus ethanol neurolysis,the VAS scores,Karnofsky scores,and PSQI scores of all the observed time points(after the operation)were statistically different compared to those before treatment and group B. Statistically difference was also observed in quality of life between two groups(P<0.05). The amount of opioid drugs of group B was statistically increased than that of group A(P<0.01).The most common side effects in Group A were postural hypotension(6 cases),diar?rhea(2 cases),and intercostal neuralgia,while nausea(20 cases),constipation(11 cases)and dizziness(8 cases)were seen in the Group B. Con?clusion Visceral nerve plexus ethanol neurolysis through post?curs of diaphragm approach by the guide of CT is effective and safe for the patients with pancreatic cancer pain,and the complications were totally acceptable.

Objective To evaluate the therapeutic effects of somatostatin(stilamin)and rhubarb for severe acute pancreatitis(SAP).Method A total of 42 patients with SAP received traditional treatment in combination with somatostatin(stilamin)and rhubarb,and compared with 40 SAP patients with routine treatment.The changes of acute physiology and chronic health evaluationⅡ(APACHEⅡ),serum amylas,serum creatinine,blood calcium,blood glucose,white blood cell count,the duration of abdominal pain,abdominal bulge,fast and hospital stay,complications,morlality and operation rate on the fist day,third day and fifth day were compared between two groups.Results Somatostatin and rhubarb reduced the complications,operation rate and mortality, and shortened the duration of abdominal pain,abdominal bulge,fast and hospital stay.Conclusions Combination of somatostatin and rhubarb is effective in the treatment of SAP patients.

This study aimed at unfolding the therapeutic effects of freeze-dried powder separated from Shuang Xia decoction (SXD) on female senile drosophila melanogaster with fragmented sleep.Taking drosophilas as the model organisms,the locomotor activity was monitored using the autonomic monitoring software to explore the regulation of fragmented sleep in senile drosophilas by the treatment of SXD.As a result,it was found that the optimum concentration of SXD was 2.50％,while the desirable therapeutic duration was 4 days.In comparison with the control group,35-day-old virgin drosophilas'sleep was prolonged in the SXD group and the positive control group.The positive drug mainly affected their sleep in the daytime,while SXD impacted their sleep at night featuring the prolonged fragmented sleep.In conclusion,it was demonstrated that the freeze-dried powder of SXD effectively alleviated intermittent insomnia in the female senile drosophilas.Compared with positive drug,SXD also mitigated intermittent insomnia at night without significant changes in the sleep of the drosophilas in the daytime.Above all,SXD was beneficial in the regulation of sleep-wake rhythm in the drosophilas.

Objective To investigate predisposing factors for early infection in patients with severe acute pancreatitis.Methods The clinical and laboratory data including age、gender、APACHE Ⅱscore on admission、hemodiastase、mechanical ventilation、blood calcium、mean arterial blood pressure、blood glucose、 alanine aminotransferase、aspartate aminotransferase、total bilirubin、necrosis of the pancreas、hypoxemia、 entero-functional disturbance、etiological factor、serum albumin、serum creatinine、urea nitrogen and haematocrit were analyzed by multiple linear regression in relation with the infection incidence in the 86 SAP patients hospitalized from Jan 2002 to Mar 2007.Results The fasting time、hiliary panereatitis、 hypoxemia、necrosis of pancreas、entero-functional disturbance、serum creatinine、urea nitrogen and haematocrit were positively correlated with the incidence of pancreatic infection(all P

OBJECTIVETo discuss arthroscopic technique of single Kirschner wire and suture avoiding epiphyseal line fixation for tibial intercondylar eminence fracture and its clinical results.

METHODSFrom May 2008 to December 2012, 21 patients (13 males, 8 females, ranging in age from 6 to 14 years old) with tibial intercondylar eminence fracture were treated arthroscopically with single Kirschner wire and suture avoiding epiphyseal line fixation technique. According to Meyers and McKeever classification, 7 patients were type II, 10 patients were type III, and 4 patients were type IV. Active rehabilitation began at one week after operation. The patients were followed up for 10 to 30 months. X-ray films were taken to evaluate fracture healing at 1, 3, 6 months after operation; range of motion, the anterior drawer test, the lachman test and the Lysholm knee score were used to evaluate clinical effects.

RESULTSAll fractures were healed without displacement at 6 weeks after operation. Anterior drawer test and the lachman test were both negative in all patients at 3 months after operation. Lysholm knee score was 95.5 ±2.5 at 6 months after operation, and postoperative X-ray film did not find epiphyseal line broadening or narrowing.

CONCLUSIONArthroscopic treatment for tibial eminence intercondylar fracture with single Kirschner wire and 8-shaped suture avoiding epiphyseal line fixation technique has many advantages, such as firm fixation,early mobilization, less invasive, less injury of physis and satisfactory effect.

Adolescent ; Arthroscopy ; Bone Wires ; Child ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Sutures ; Tibial Fractures ; surgery

Soil salinity is an important issue, as most crop plants are low in salt tolerance. Salt tolerance, a complex, multifactorial, and multigenic process, has been known to be a quantitative trait. The identification of the salt stress responsive genes or salt tolerance genes is essential for the breeding programs. Most recent efforts have been focused on the products of structural genes (transport proteins, ion channels, enzymes of solute synthesis) while little attention were paid to the regulatory aspects of these proteins. Since the first aquaporin gene from plants was cloned and functionally expressed in 1993, there has been a growing interest in the molecular biology of MIPs (membrane intrinsic proteins) and their bearing on the biophysics of water flow across plant membranes. In the last decades, studies on Mangroves, a special kind of wood plants, grow in high-salt and flooding conditions have been concentrated almost exclusively on their physiological and ecological characteristics. Kandelia candel, one of the dominant species of mangroves along the Chinese coast, lacks salt glands or salt hairs used for removal of excess salt in other mangroves. This makes K. candel a perfect model to study the molecular mechanism of salt tolerance in mangrove plants. Using cDNA RDA, a cDNA-specific modification of genomic representational difference analysis, a series of salt responsive genes of Kandelia candel were cloned. Among these gene fragments, a 183 bp fragment (termed as SRGKC1) encoding a tonoplast intrinsic protein (TIP) in Kandelia candel (KCTIP1) was identified. Based on the sequence of SRGKC1, two gene specific primers were designed, and the 3' and 5' end of the KCTIP1 gene were obtained using the SMART RACE cDNA Amplification Kit. RACE products were purified from low-melting agarose, and sequenced directly with GSPs as the sequencing primers. A 500-bp fragment corresponding to the 3'end of this gene was obtained using the GSP1 primer, and a 690 bp fragment corresponding to the 5' end of this gene was obtained using the GSP2 primer. Two primers that flank the putative open reading frame (ORF) were designed to obtain the cDNA containing the complete ORF by RACE PCR reaction. The full-length cDNA of KCTIP1, containing a 756 bp open reading frame (ORF), was approximately 1.1 kb; the start codon was located at the nucleotides of 99-101 and stop codon at the nucleotides of 855-857 followed by a poly (A) tail. The KCTIP1 cDNA sequence in this research was released in GenBank with accession number AF521135. Using ExPASy Proteomics tools provided by EMBL, the isoelectric point and MWt of KCTIP1 are estimated as 5.77 and 26.3 kD respectively. Transmembrane prediction analysis revealed the deduced KCTIP1 protein sequence contains six transmembrane regions at amino acid residues of 20 - 42, 57 - 79, 86 - 108, 113 - 135, 142 - 164 and 217 - 239. Two highly conserved asparagine-proline-alanine (NPA) motifs were located at 85 - 87 and 199 - 201 amino acid residues respectively. KCTIP1 is also predicted to contain the Cys residue (Cys 118) that are shown to confer Hg-sensitivity in Arabidopsis gamma-TIP and delta-TIP. Similarity analysis showed that KCTIP1 shared 77% - 79% amino acid sequence identity with the TIPs from Vitis berlandieri, Brassica oleracea and Arabidopsis thaliana. Expression analyses indicated that KCTIP1 had different expression among species of Mangroves. Expressions of KCTIP1 in Kandelia candel, Rhizophora apoculata and Ceriops tagal were suppressed by salt, and were insensitive to salt stress in unknown species of Mangroves. Previous studied showed that salt conditions might result in large and rapid changes in extracellular water potential and serious disturbance to the cytoplasm. In order to compensate for this imbalance, the relative contribution of water channels to flow across the root could thus vary. K. candel is a species that is native to intertial zone of tropical and subtropical coast and is well-adapted to salt conditions. The coordinated down-regulation of aquaporins in this plant may decrease membrane water permeability and thus increase the cellular water conserva- tion during periods of salt stress. The results reported here are consistent with the postulated roles for tonoplast water channels in regulating the hydraulic permeability of the vacuolar membranes and in adjusting the water homeostasis of the protoplasm under various physiological conditions. The identification of KCTIP1 as one of salt-responsive genes implies that intracellular osmotic equilibration is a part of salt-tolerant mechanisms in Mangroves.
Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; DNA, Complementary ; genetics ; Membrane Proteins ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Plant Proteins ; chemistry ; genetics ; Rhizophoraceae ; genetics ; Sequence Alignment

OBJECTIVETo investigate the effect of peroxiredoxin I (Prx I) gene silencing on the radiosensitivity of breast carcinoma MCF-7 cell xenograft in nude mice and explore the mechanism.

METHODSMCF-7 cells were transfected with the recombinant plasmids pGPU6-PrxI and pGPU6-HK separately. The pGPU6-PrxI-transfected cells stably expressing Prx I shRNA and pGPU6-HK-transfected cells were inoculated subcutaneously into BALB/c nude mice. After exposure to ionizing radiation (IR) with 6 MV X-ray, the xenografts were harvested for measuring the tumor volume and mass, and the tumor inhibition rates were calculated. Immunohistochemistry was employed for detecting the expressions of Prx I and caspase-3 proteins. The ultrastructural changes of the tumor tissues following the exposure were observed using electron microscopy. Western blotting was used to analyze the expressions of γ-H2AX and Rad51 proteins.

RESULTSFollowing IR exposure, the pGPU6-Prx I-transfected cell xenograft showed a significantly delayed growth and smaller tumor volume as compared with pGPU6-HK xnegraft, with a tumor inhibition rate reaching 79.76%, significantly higher than that in non-exposed pGPU6-Prx I group (34.92%) and pGPU6-HK+IR group (56.94%) (P<0.05). The pGPU6-Prx I-transfected xenografts showed significantly increased tumor cell apoptosis and necrosis, down-regulated the expressions of Prx I and Rad51 proteins, and up-regulated the expressions of caspase-3 and γ-H2AX proteins; these changes were even more obvious after IR exposure, which caused a decrease of Rad51 protein by 84.8% and an increase in γ-H2AX protein by 5.6 folds compared with those in pGPU6-HK group (P<0.05).

CONCLUSIONPrx I gene silencing can significantly enhance the radiosensitivity of breast carcinoma xenograft in nude mice possibly by increasing DNA damage and lowering the capacity of the cells for DNA repair. Prx I may serve as an ideal molecular target for radiosensitization of breast carcinoma.

Animals ; Breast Neoplasms ; genetics ; pathology ; radiotherapy ; DNA Repair ; genetics ; Female ; Humans ; MCF-7 Cells ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Peroxiredoxins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Radiation Tolerance ; genetics ; Transfection ; Xenograft Model Antitumor Assays