Overexpression of procollagen gene can cause the extraordinary increase of collagen's synthesis and therefore lead to the keloid and hypertrophic scar. To utilize ribozyme to suppress the expression of procollagen genes, a eukaryotic expression recombinant plasmid containing a dual-ribozyme gene against alpha 1 (I) and alpha 1 (III) procollagen genes was constructed. The ribozyme from in vitro transcription was incubated with target transcripts from recombinant plasmids which separately contained the fragments of the second exons of pro alpha 1 (I) and pro alpha 1 (III) collagen genes under various experimental conditions. The results showed that the dual-ribozyme could efficiently catalyze the specific cleavage of the target RNAs at 37 degrees C, 42 degrees C, 50 degrees C and Mg2+ concentration from 10 mmol/L to 20 mmol/L. This work provided a basis for further study on the ribozyme to suppress the expression of procollagen genes and control the cicatrization.
Base Sequence ; Exons ; Molecular Sequence Data ; Procollagen ; genetics ; RNA ; metabolism ; RNA, Catalytic ; genetics ; metabolism ; Temperature

The dense extracellular matrix (ECM) of the tumor severely limits the deep penetration of nanomedicine and weakens its anti-tumor effect. Based on this, the yeast vesicle biomimetic nanomedicine with active deep penetration ability of tumor tissue was designed and developed for enhanced tumor therapy. Results of characterization showed that the yeast cell vesicles (YCV) displayed a spherical morphology with diameter of around 100 nm and was well dispersed. Then the chemotherapeutic drug doxorubicin (DOX) was selected as a model drug, and DOX was loaded into YCV to obtain YCV/DOX through electrostatic interaction, the encapsulation efficiencies of DOX were calculated as 82.5%. The drug release profile of YCV/DOX implied that DOX release showed a manner of pH-dependent, it may be that pH has affected the electrostatic effect of YCV and DOX. Compared with liposomes (Lipo), in vitro cell experiments showed that YCV from natural sources had stronger permeability in three-dimensional multicellular spheres. It is speculated that the mechanism may be good deformation capacity of YCV. A 4T1 xenograft tumor model was established to evaluate the therapeutic efficacy of YCV/DOX. The results suggested that YCV/DOX has stronger tumor tissue penetration ability and could effectively inhibit the tumor growth. All animal experiments were performed in line with national regulations and approved by the Animal Experiments Ethical Committee of Zhengzhou University. This study brings new ideas for the development of biomimetic nanomedicine to overcome the ECM of solid tumors.

OBJECTIVETo explore the pathoclinical features of adult primary mediastinal liposarcoma and their correlation with the prognosis.

METHODSThe clinical data of 19 patients with adult primary mediastinal liposarcoma who were treated in our hospital between 1970 and 2011 were retrospectively analyzed. Eighteen patients underwent open thoracic tumor excisions for at least one time, and the remaining one patient only received biopsy surgery. Histopathological results after surgery revealed that 6 well-differentiated type tumors, 6 myxoid type tumors, 3 pleomorphic type tumors, 2 mixed type tumors, and 2 un-classified tumors.

RESULTSAmong 6 patients with well-differentiated type tumors, 1 died after having been survived for 10 years; 5 were still alive, with a mean duration of 126.2 months,the 5-year survival rate was 100%. Among 6 patients with myxoid type tumors, 5 patients had follow-up data, with a mean survival of 26.2 months. Among 3 patients with pleomorphic type tumors, only one patient had follow-up data: the patient finally died, with a survival of 34 months. Of 2 patients with mixed type tumors, only one patient had follow-up data: the patient survived 8 months and died. Of 2 patients with un-classified type tumors, one had follow-up data: the patient lived for 24 months and died.

CONCLUSIONSMediastinal liposarcoma is a rare disease. Surgery is the primary therapeutic modality. Different pathological subtypes have different epidemiological features, biological behaviors, and malignant potentials. Pathological subtype is an important prognosis factor. Patients with well-differentiated tumors have much better prognosis than those with other subtypes.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Liposarcoma ; pathology ; surgery ; Male ; Mediastinal Neoplasms ; pathology ; surgery ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo assess the clinical outcomes of brachiocephalic artery-sparing aortic arch repair combined with stent-graft elephant trunk technique for treatment of Stanford type A aortic dissection.

METHODSTwenty-three patients with Stanford type A aortic dissection requiring arch replacement underwent brachiocephalic artery-sparing aortic arch repair combined with stent-graft elephant trunk technique. The operations were performed within 72 h (20 cases) or 3-14 days (3 cases) after the onset of aortic dissection.

RESULTSThere was no perioperative death in these cases. The mean extracorporeal circulation time was 209∓52 min, the aortic cross clamp time was 85∓21 min, and the mean chest tube output within the first 24 h after the operation was 570∓263 mL; none of the patients required chest reopening for management of bleeding. Postoperative acute renal failure requiring hemodialysis occurred in 3 cases, transient neurologic dysfunction in 2 cases, paraplegia in case and hematosepsis in 1 case. No such complications as permanent neurologic deficit or postoperative visceral malperfusion occurred in these cases. All the patients survived and were discharged from hospital without experiencing severe complications in the follow-up for 6-18 months.

CONCLUSIONBrachiocephalic artery-sparing aortic arch repair combined with stent-graft elephant trunk technique is a safe and simple procedure with controllable bleeding and can serve as an optional procedure for aortic arch replacement.

OBJECTIVETo compare the therapeutic effects and safety of herpes zoster treated by the cotton sheet moxibustion combined with the plum-blossom-needle tapping therapy to western medicine.

METHODSThe multicentral random controlled method was adopted, 120 cases of herpes zoster were randomly divided into a comprehensive treatment group and a western medication group, 60 cases in each one. In the comprehensive treatment group, the tapping therapy of plum blossom needle was applied to the foci, corresponding Jiaji (EX-B 2), Quchi (LI 11), Waiguan (TE 5), Zusanli (ST 36), Taichong (LR 3), etc. Afterward, the cotton sheet moxibustion was given. In western medication group, Acyclovir ointment for external application, Valaciclovir Hydrochloride tablets and Vitamin B1 for oral administration were prescribed. In 7 days of treatment, the clinical symptom score, effect time, efficacy and safety were observed before and after treatment between two groups. The recurrence of disease was followed up for 1 month.

RESULTSIn the comprehensive treatment group, the cured rate and the total effective rate were 80.0% (48/60) and 98.3% (59/60) separately, which were significantly better than 45.0% (27/60) and 71.7% (43/60) in western medication group separately (P < 0.01, P < 0.05). After treatment, in either group, the scores of clinical symptoms such as pain rating index (PRI), Visual Analogue Scale (VAS), present pain intensity (PPI), skin lesion and sleeping score, etc. were all reduced significantly (P < 0.01, P < 0.05). The score reducing was much more obvious in the comprehensive treatment group (P < 0.01, P < 0.05). In the comprehensive treatment group, the time of pain stopping, the time of blister stopping, the time of scarring and the time of healing were all shorter tha tn those in western medication group (P < 0.01, P < 0.05). In the follow-up observation, 1 case (1.6%) was recurred in the comprehensive treatment group and 8 cases (13.3%) were in western medication group. In western medication group, 6 cases presented mild adverse reactions.

CONCLUSIONThe cotton sheet moxibustion combined with the plum-blossom-needle tapping therapy is advantageous at good efficacy, quick effect and short-time treatment for herpes zoster, which is apparently superior to the treatment with Acyclovir ointment for external application, Valaciclovir Hydrochloride tablets and Vitamin B1 for oral administration. It is the safe and effective therapy.

Adult ; Aged ; Female ; Herpes Zoster ; therapy ; Humans ; Male ; Middle Aged ; Moxibustion ; Pain Management ; Young Adult

OBJECTIVETo prepare ligustrazine (TMPZ) ocular sustained-release implant, and investigate its in vitro drug release, pharmacokinetics in rabbit vitreum and in vitro correlation.

METHODLigustrazine ocular sustained-release implants were prepared by micro-twin conical screw mixers with hot-melting extrusion method, with polyactic-co-glycolic acid (PLGA) as the matrix. HPLC was adopted to determine the concentration in vitreum after ligustrazine was implanted in rabbit eyes, in order to examine its in vivo sustained-release behavior, and study the correlation between in vitro and in vivo.

RESULTLigustrazine implants were prepared with a drug-loading rate between 10% and 30%, which was in conformity to the pharmacopoeia in terms of the content uniformity. Its in vitro release was in conformity to the zero-order release model. With PLGA 5050, 2. 5A as a vector, ligustrazine implants with a drug-loading rate of 30% could slowly release drug for more than 3 weeks, indicating a good correlation between in vitro and in vivo release.

CONCLUSIONLigustrazine ocular implants prepared with hot-melting extrusion method is practicable. Ligustrazine ocular implants release drug smoothly in rabbit vitreous vitreums, suggesting good sustained-release effect.

Animals ; Biological Availability ; Drug Implants ; Eye ; Female ; Male ; Polyglycolic Acid ; chemistry ; Pyrazines ; administration & dosage ; chemistry ; pharmacokinetics ; Rabbits ; Vitreous Body

OBJECTIVETo construct a Gpx1 and klk1 recombinant vector containing the kidney-specific promoter Ksp-cadherin.

METHODSHuman Gpx1, Klk1 and Ksp-cadherin cDNAs were amplified with PCR and inserted in a stepwise manner into the expressive vector pIRES-EGFP to construct the recombinant vector Ksp-cadherin-Gpx1-Klk1. The constructed vector was verified with restriction enzyme digestion and sequence analysis.

RESULTS AND CONCLUSIONThe recombinant expression vector Ksp-cadherin-Gpx1-Klk1 was constructed and identified successfully, which provides a potent tool for preparing transgenic animals to investigate gene therapy for ischemia-reperfusion injury in kidney transplantation.

Cadherins ; genetics ; Cloning, Molecular ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Glutathione Peroxidase ; genetics ; Humans ; Kallikreins ; genetics ; Kidney ; metabolism ; Promoter Regions, Genetic ; genetics

The proteins encoded by the Brassica juncea chitinase gene BjCHI1 and its derived genes BjCHI2 and BjCHI3 were expressed by Multi-copy Pichia expression system. The chitinase activity of FPLC purified BjCHI1, BjCHI2 and BjCHI3 were tested and the results showed that all the three proteins degraded both CM-chitin-RBV and colloidal chitin. The Km values of BjCHI1, BjCHI2 and BjCHI3 for CM-chitin-RBV were estimated as 0.799 mg/mL, 0.544 mg/mL and 0.793 mg/mL, respectively. When the colloidal chitin was used as substrate, the Km values were 0.281 mg/mL, 0.388 mg/mL and 1.643 mg/mL, respectively, indicating chitin-binding domain can increase affinity of chitinase to insoluble substrate. In the agglutination activity assay, only BjCHI1 shows activity when the protein concentration was more than 33 micrograms/mL, while BjCHI2 and BjCHI3 without agglutination activity even when the concentration was increased as high as 800 micrograms/mL. This means that the two chitin-binding domains in BjCHI1 are essential for agglutination and BjCHI1 is the first protein which shows both chitinase and agglutination activity identified so far in plants.
Agglutination ; Agglutinins ; genetics ; Chitinases ; genetics ; isolation & purification ; physiology ; Mustard Plant ; chemistry ; Pichia ; genetics ; Plant Proteins ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; isolation & purification

Diacylglycerol, DAG, because of its multifunctional and nutritional properties, attracted considerable attention recently. Enzymatic synthesis of diacylglycerols from linoleic acid was investigated in a solvent-free reaction in a continuously operated fixed bed reactors containing Lipozyme RM IM. By appropriate manipulation of the fluid-residence time, the relative proportions of the various acylglycerols in the effluent stream can be controlled. In addition, the presence of excess glycerol is effective for the removal of water produced during the esterification reactions. Under the conditions of molar ratio of linoleic acid to glycerol of 0.5, the immoblized enzyme maintained high stability and allowed the reaction to continue for 10 days without significant deterioration in enzyme activity. It was determined that the conversion of fatty acid, content of 1,3-DAG and volume efficiency of reactor reached optima under the conditions: a packaged-bed reactor(with a ratio of packed length to inner diameter of 7.8), reacting temperature at 65 degrees C, molar ratio of linoleic acid to glycerol of 0.5, and feeding flow rate of 1.2 mL/min.
Catalysis ; Diglycerides ; chemical synthesis ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Lipase ; chemistry