Adolescent ; Adult ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; blood ; Humans ; Male ; Middle Aged ; Norepinephrine ; blood ; Young Adult

OBJECTIVEBy using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target.

METHODSDesigned siRNA aims at Itk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory cytokines by ELISA.

RESULTSItk protein can be suppressed by Itk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-gamma. They all have statistical difference (P < 0.05).

CONCLUSIONItk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokines.This can supply an experimental basis to regard Itk as drug target for inflammation therapy.

Animals ; Cell Differentiation ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein-Tyrosine Kinases ; genetics ; immunology ; Spleen ; cytology ; immunology

OBJECTIVETo explore the clinical outcomes of open reduction and internal fixation with calcaneal locking plates in treating Sanders type II and III calcaneal fractures.

METHODSFrom January 2010 and October 2012, 38 calcaneal fractures with Sanders type II or III were treated with open reduction and internal fixation with calcaneal locking plate. According to the Sanders classification, 15 fractures were classified as type II, 23 fractures as type III. The patients were divided into two groups (group A and B) according to the different fixed methods. Sustentaculum tali was fixed with one screw in group A, including 13 males and 5 females, with a mean age of (38.56±8.03) years old (ranged, 25 to 55). And sustentaculum tali was not fixed in group B, including 16 males and 4 females, with a mean age of (42.35±8.29) years old (ranged, 29 to 53). Clinical effects were evaluated according to the changes of Böhler's angle and the Maryland Foot Score and VAS score.

RESULTSAll patients were followed up from 12 to 20 months with a mean of 14 months. Böhler's angles and subtalar joints obtained satisfactory reconstruction in all patients. One year after operation, the mean Maryland Foot Score was 88.61±7.59 in group A; and was 82.40±9.24 in group B; Maryland Foot Score of group A was higher and foot functional rehabilitation was better than group B. The mean VAS score was 13.39±11.47 in group A; and was 22.50±13.10 in group B; VAS score of group A was lower and foot pain was less than group B.

CONCLUSIONSustentaculum tall screw fixation has advantages of strong fixed strength, high stability, less postoperative pain, rapid functional recovery in treating Sanders type II and III calcaneal fractures.

Adult ; Bone Plates ; Bone Screws ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function

Objective To establish a new determination method for the measuring of alkaline phosphatase activity (ALP) with p-acetyl phenyl phosphace (PAP-PNa_2) as substrate.Methods With the help of Vital semiautomatic analyzer,researched a continuous-monitoring procedure and set up experimental parameters.Results When using this assay,the wavelength of PAP's absorption was 325 nm and the Km of ALP was 0.376 mmol/L.The molecular extinction coefficient of PAP at 340 nm was 23 390 L?mol~(-1)? cm~(-1) and the concentration of citrate buffer was 0.438 mol/L.During the process,we found that the optimum pH of enzyme was 10.4,and the concentration of substrate was 5.0 mmol/L.The time of linear reaction was 900 seconds,and the linear range was 0-1 110 U/L.Serum total ALP were 63.1-118.3 U/ L(male) and 52.5-89.0 U/L(female),based on results from 60 heath adults.Conclusions The method is practical in its repetition and convenience,saves time and is not liable to be affected by bilirubin in serum.It is especially suited to the use of automatic analyzers.

This study is to explore new lead compounds by inhibition of Pin1 for anticancer therapy using temperature sensitive mutants. As Pin1 is conserved from yeast to human, we established a high-throughput screening method for Pin1 inhibitors, which employed yeast assay. This method led to the identification of one potent hits, 8-11. In vitro, 8-11 inhibited purified Pin1 enzyme activity with IC50 of (10.40 +/- 1.68) micromol x L(-1), induced G1 phase arrest and apoptosis, showed inhibitory effects on a series of cancer cell proliferation, reduced Cyclin D1 expression, was defined as reciprocally matched for protein-ligand complex in virtual docking analysis and reduced cell migration ability. In vivo, we could observe reduction of tumor volume after treatment with 8-11 in xenograft mice compared with vehicle DMSO treatment. Altogether, these results provide for the first time the involvement of 8-11 in the anticancer activity against Pin1.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Drug Screening Assays, Antitumor ; methods ; G1 Phase ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; NIMA-Interacting Peptidylprolyl Isomerase ; Neoplasms ; pathology ; Peptidylprolyl Isomerase ; antagonists & inhibitors ; Temperature ; Xenograft Model Antitumor Assays ; Yeasts

OBJECTIVETo evaluate the possibility of short interfering RNA (siRNA) inhibiting Coxsackievirus B3 (CVB3) infection in vitro, and discover the mechanism initially.

METHODSWe obtained proper effective dosage of siRNA by observing cytopathic effect (CPE). Estimate its antiviral activities and its pathway of siRNA by Western Blot assay and RT-PCR.

RESULTSResults showed that siRNA-3753 can be effectively transfected into HeLa cells, we can achieve a high transfection efficiency up to 98.77% and its effect can last for 48 h stably in cells. 0.6 micromol/L siRNA-3753 got a high inhibiting effect of virus and didn't show any toxicity to cells. So we consider this concentration as the experimental concentration. siRNA-3753 can debase virus reproduction. The antiviral effect is sequence-specific and is not attributable to either interferon or the interferon response effectors protein kinase R (PKR).

CONCLUSIONThe data confirmed that siRNA can effectively inhibit CVB3 infection in vitro, its antivirus effect was gained from specific debase of virus genome.

Coxsackievirus Infections ; therapy ; virology ; Enterovirus B, Human ; genetics ; metabolism ; HeLa Cells ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; RNA, Viral ; genetics

Objective To examine the expression of human ?-defensin 2 (hBD_ 2 ) recombinant adenovirus expression vector in rat dermal multipotent stem cells (dMSCs) and to observe the antiseptic activity of recombinant hBD_ 2 . Methods The expression of hBD_ 2 in dMSCs was examined by RT-PR, fluorescent immunochemistry and Western blotting, and the concentration of recombinant hBD_ 2 in supernate was measured by ELISA. The antiseptic activity of recombinant hBD_ 2 was assessed by K-B disc agar diffusion test. Results hBD_ 2 could be effectively expressed in dMSCs, and the concentration of recombinant hBD_ 2 in supernate was about 743.6 ng/ml . Recombinant hBD_ 2 in supernate showed antiseptic activity. Conclusion Recombinant adenovirus expression vector of hBD_ 2 could be effectively expressed in dMSCs, and the recombinant hBD_ 2 in supernate showed obvious antiseptic effects toward some standard bacteria lines.

Adult ; Disease Outbreaks ; Humans ; Metapneumovirus ; Paramyxoviridae Infections ; epidemiology ; Respiratory Tract Infections ; epidemiology ; virology ; Students

To evaluate the effects of lentivirus-delivered short hairpin RNA (shRNA) on CVB3 infection in an animal model by RNA interference technique, we constructed a recombinant lentivirus expressing shRNA-3753 against the viral genome region 3753-3771, then transduced Lenti-sh3753 into mice infected with CVB3. We evaluated the antiviral ability of lenti-sh3753 by cytopathic effect (CPE), viral plaque assay and histological analysis of mice hearts. The results showed that Lenti-sh3753 exhibited a significant protective effect on cell viability and reduction of viral titers in supernatant of cell culture by specific inhibition on viral replication. Lenti-sh3753 also prolonged the mice survival and limited the viral production in mice hearts. These data proposed that Lenti-sh3753 can effectively inhibit CVB3 infection in a coxsackievirus-induced myocarditis model, suggesting its potential role in prevention and therapy of viral diseases.
Animals ; Coxsackievirus Infections ; drug therapy ; virology ; Down-Regulation ; Enterovirus B, Human ; genetics ; physiology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; drug therapy ; virology ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; RNA, Viral ; genetics ; Virus Replication

Acremonium ; chemistry ; Adjuvants, Immunologic ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Cordyceps ; chemistry ; classification ; Humans ; Hypoglycemic Agents ; pharmacology ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology