Antiviral Agents ; therapeutic use ; Hepatitis C ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; virology

To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Humans ; Oligopeptides ; biosynthesis ; pharmacology ; Pichia ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; pharmacology

China ; epidemiology ; Data Collection ; standards ; Humans ; Occupational Diseases ; chemically induced ; epidemiology ; Occupational Exposure ; statistics & numerical data ; Occupational Health ; statistics & numerical data ; Occupational Medicine ; legislation & jurisprudence

Objective To explore the effects of the therapeutic effects of interventional heated chemotherapy( IHC) for moderate and advanced huge primary hepatocellular carcinoma and compare with transcatheter arterial chemoembolization( TACE) . Methods The 68 pa-tients with moderate and advanced huge primary hepatocellular carcinoma accepted the treatment of IHC were considered as the observation group,and 32 patients managed TACE were regarded as the control group. The therapeutic effects,living quality,secondary surgery rates,The reduced level of serum AFP,survival rate,and toxic side effects of 2 groups were contrasted. Results The effective rate of observation group was obviously higher than that of control group (79. 4% vs 56. 3%,P <0. 05),and secondary surgery rates was increased(70. 6% vs 43. 8%,P<0. 01). The ratio of increased Karnofsky score was more than 20 in observation group,which was obviously higher than that of control group(35. 3% vs 12. 5%,P<0. 01). T The reduced level of serum AFP was more than 50% in observation group,which was higher than that of control group (52. 9% vs 25. 0%,P<0. 01). The survival rate of 12,18,24 months after treatment in control group was in-creased than that in control group individually (P<0. 01). Conclusion IHC is effective for huge primary hepatocellular carcinoma and in-creases the secondary surgery rates,which is one of the safe and effective methods for huge primary hepatocellular carcinoma.

One of the key contents in tissue engineering trachea replacement research is the scaffold selection.This review summarizes the latest original literatures and investigations about electrospun technique as well as recent progress.To discuss the advantages and disadvantages of natural,synthetic and combined electrospun scaffolds,the versatility in material choosing and production methods is the unique superiority.For specific experimental or clinical objects,the further research is to choose a suitable polymer,to improve surface modification techniques and to control the dimension and arrangement of the fibrous structure of electrospun fibers,which can offer versatility and tissue specificity,and therefore provide a bright prospect for tissue engineering trachea.

Bile Duct Neoplasms ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; pathology ; surgery ; Humans

Animals ; Diet, High-Fat ; Glucagon ; blood ; Glucose ; metabolism ; Hormones ; blood ; Insulin ; blood ; Leptin ; blood ; Lipid Metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Animals ; Hypertension ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism

Objective:To construct the yeast expressive vector of rmFⅦ, in which mFⅦ was mutated to inhibit coagulation without affecting the affinity for TF, and express it in Pichia pastoris. Methods:The full length cDNA encoding mFⅦ was amplified from a mouse liver by RT-PCR method, site-direct mutated and restriction enzyme digested as design. Cloning into pPIC9K, electroporation of Gs115, in vivo screen of multiple inserts by G418 resistance, BMGY/BMMY are used for induction and expression of rmFⅦ in pichia pastoris. These proteins were also screened for functional activity. Results: Three different rmFⅦ-pPIC9K yeast expression vectors and it's aim protein were obtained,two kinds of proteins were found to be functional active as design. Conclusion:rmFⅦ protein can be expressed in pichia pastoris and it might facilitate the development of tumor-target molecule, and novel anti-agiogenesis drug study.

Objective To examine clinical features, neuroimaging presentation and treatment of intracranial fungal granulomas (ICFG) in order to improve the accuracy rates of diagnosis and cure. Methods Three pathologically proven cases with ICFG were retrospectively analyzed. Cases of ICFG reported in literature were also summarized. Results Among the 3 patients with ICFG, 1 patient had a history of head trauma and craniotomy and 1 had a history of type 2 diabetes mellitus and a long history of exposure to doves. In all 3 patients, the symptoms started with headache and vomiting, accompanied by low-grade fever, convulsion, and cranial nerve deficits. Intracranial mass lesion was revealed on brain computed tomography (CT) scan and (or) magnetic resonance imaging (MRI) with or without intravenous contrast. CT scan showed low-density lesions and granulations with ring and homogenous enhancement, indicating fungal abscesses. MRI in all 3 cases showed one or multiple circumscribed intracranial space-occupying lesion, with ring, heterogeneous contrast enhancement and perilesional edema. The treatments were initiated with craniotomy and surgical resection of granulations followed by intravenous injection of amphotericin B (AMB) combined with fluconazole. The daily administration of AMB was increased gradually from 0.25-1.50 mg/kg and the total dosage of 2-4 g should be achieved within 3 months. The combination therapy with fluconazole (400 mg/d ) was also given by intravenous injection. To increase penetration into cerebrospinal fluid, intrathecal injection of AMB had also been given at the maximum dosage of 1 mg every time, twice a week. Two patients were administered fluconazole (200 mg/d ) orally for 3, 6 months consecutively after completing the combination therapy of AMB with fluconazole, while the other patient refused continuous antifungal treatment 1 month postoperatively. All 3 patients were followed up for a period between 3, 24, 48 months. The 2 patients that completed full antifungal treatment were cured without recurrence. The other patient had improved transiently after operation but died after 3 months. Conclusions Because no distinct chnical and neuroimaging features are presented in ICFG, it is difficult to diagnose preoperatively. Indications for surgery include diagnosis, relief mass effect and increase efficiency of drug treatment. Use of appropriate and completed antifungal treatment decreases mortality. The treatment requires continued and long-term administration of antifungal medication to prevent relapses, whether granulomas are totally removed or not.