Altitude ; Animals ; Arvicolinae ; Gene Expression ; Hypoxia ; Rats ; Tumor Suppressor Protein p53 ; genetics

To explore the physiological role and biocatalytic properties of short-chain dehydrogenases from Pseudomonas fluorescens GIM1.49, we cloned the structural gene pfd and characterized its over-expressed product. The length of gene pfd was 684 bp encoding a short-chain dehydrogenase with 227 amino acid residues and calculated molecular mass of 24.2 kDa. The recombinant plasmid pET28b-pfd was constructed and functionally expressed in Escherichia coli BL21(DE3), resulting in the over-production of recombinant short-chain dehydrogenase PFD with a size of 28 kDa. The enzyme could oxidize alcohols including 4-chloro-3-hydroxbutanoate ester and reduce 4-chloro-acetoacetate ester using either NAD(H) or NADP(H) as coenzyme. The enzyme showed the highest activity against 4-chloro-3-hydroxbutanoate ester as substrate, with Km of 186.40 mmol/L and Vmax of 89.56 U/mg. When catalying the oxidative reaction, its optimal temperature was 12 degrees C and optimal pH was 10.5, in contrast to the values of 24 degrees C and pH 8.8 in the reductive reaction. The enzyme had high solvent tolerance and its activity was improved by the addition of Ca2+ (1 mmol/L) or EDTA (5 mmol/L). These results indicated that the enzyme from Pseudomonas fluorescens GIM1.49 was a novel short-chain dehydrogenase and might play a role in oxidative degradation of halogenated secondary alcohols.
Alcohols ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Butyryl-CoA Dehydrogenase ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Oxidation-Reduction ; Pseudomonas fluorescens ; enzymology ; genetics ; Recombinant Proteins ; genetics ; metabolism

MicroRNA (miRNA) is an endogenous, non-coding single-stranded RNA that regulates a variety of signal pathways or cytokines.Recent studies have confirmed that miRNA can affect alkaline phosphatase activity and matrix mineralization in bone formation, and plays an important role in osteogenic differentiation and cartilage differentiation.Abnormalities in the osteogenesis process can lead to osteogenesis imperfecta, Feingold syndrome, and femoral head necrosis.This review summarizes the specific mechanism of osteogenic differentiation and cartilage differentiation regulated by miRNA, suggesting the new clue for the future research about the underlying mechanism of bone development and clinical treatment of bone dysplasia from epigenetics.

Objective To compare the clinical effects of microneurosurgery by supraorbital key-hole or endonasal transsphenoidal approaches in the treatment of pituitary adenoma and investigate their complications. Methods We retrospectively analyzed the data of 87 patients with pituitary adenoma of which the anteroposterior diameter was less than 3 cm. These patients, admitted to our hospital from May, 2006 to June, 2008, were operated in an endoscope-assisted microsurgical manner via a supraorbital key-hole approach (n=42) or an endonasal transsphenoidal approach (n=45). The efficacy of these two approaches was compared and their complications were observed. Results The excision rate of the pituitary adenoma developing on or around the sella turcica operated through the supraorbital key-hole approach was significantly higher than that through the endonasal transsphenoidal approach(P< 0.05); while that of microadenoma or adenoma developing towards the sphenoid sinus operated through the supraorbital keyhole approach was statistically lower than that through the endonasal transsphenoidal approach (P<0.05). No obvious differences on the improvement of endocrine secretion, visual acuity and field was noted in these two approaches (P>0.05). The incidence rate of epistaxis and unilateral dysosphresia in the supraorbital key-hole approach was significantly lower as compared with that in the endonasal transsphenoidal approach (P<0.05). Conclusion Rarely having such complications as dysosphresia, epistaxis and sphenoiditis, neuroendoscopic surgery through supraorbital key-hole approach is the best way of treating the pituitary adenoma developing on or around the sella turcica and worth to promote in clinic.

Adult ; Aged ; Antigens, CD ; biosynthesis ; genetics ; B7-H1 Antigen ; CD28 Antigens ; biosynthesis ; genetics ; Female ; Hepatitis B, Chronic ; immunology ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Middle Aged

OBJECTIVEMalignant middle cerebral artery (MCA) infarction is characterized by mortality rate of up to 80%. The aim of this study was to determine the value of decompressive craniectomy in patients presenting malignant MCA infarction compared with those receiving medical treatment alone.

METHODSPatients with malignant MCA infarction treated in our hospital between January 1996 and March 2004 were included in this retrospective analysis. The National Institute of Health Stroke Scale (NIHSS) was used to assess neurological status on admission and at one week after surgery. All patients were followed up for assessment of functional outcome by the Barthel index (BI) and modified Rankin Scale (RS) at 3 months after infarction.

RESULTSTen out of 24 patients underwent decompressive craniectomy. The mean interval between stroke onset and surgery was 62.10 h. The mortality was 10.0% compared with 64.2% in patients who received medical treatment alone (P<0.001). The mean NIHSS score before surgery was 26.0 and 15.4 after surgery (P<0.001). At follow up, patients who underwent surgery had significantly better outcome with mean BI of 53.3, RS of 3.3 as compared to only 16.0 and 4.60 in medically treated patients. Speech function also improved in patients with dominant hemispherical infarction.

CONCLUSIONDecompressive craniectomy in patients with malignant MCA infarction improves both survival rates and functional outcomes compared with medical treatment alone. A randomized controlled trial is required to substantiate those findings.

Adult ; Aged ; Craniotomy ; methods ; Decompression, Surgical ; methods ; Female ; Humans ; Infarction, Middle Cerebral Artery ; diagnosis ; surgery ; Male ; Middle Aged ; Recovery of Function ; Retrospective Studies ; Severity of Illness Index ; Treatment Outcome

OBJECTIVETo observe the effectiveness and safety of electroacupuncture (EA) at Neimadian (Extra) and Neiguan (PC 6) for analgesia after thoracic surgery.

METHODSOne hundred and twenty cases of thoracic surgery were randomly divided into an electroacupuncture (EA) group (60 cases) and a medication group (60 cases). EA was applied at Neimadian (Extra) and Neiguan (PC 6) for postoperation analgesia in the EA group, while patient-controlled intravenous analgesia (PCIA) was applied in the medication group. The score of visual analogue scale (VAS), analgesia effect, safety and beta-endorphin level after the treatment in both groups were compared.

RESULTSCompared with those before the treatment, the VAS scores in every time point after surgery were decreased (all P < 0.05), which were lower in the EA group (P < 0.01). The excellent and good rates were 96.7% (58/60) and 75.0% (45/60) seperately, the analgesia effect in the EA group (2 h after operation) was superior to that in the medication group (P < 0.01). The safety degree in EA group was higher to that in the medication group (P < 0.01). Compared with that before the treatment, the beta-endorphin level in two groups after treatment was both increased, which was higher in the EA group (P < 0.01).

CONCLUSIONElectroacupuncture at Neimadian (Extra) and Neiguan (PC 6) has better analgesia effect (2 h after operation) and safety than PICA on analgesia after thoracic surgery.

Acupuncture Analgesia ; Acupuncture Points ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; blood ; therapy ; Thoracic Surgery ; beta-Endorphin ; blood

OBJECTIVETo explore the applied feasibility of the anterior cervical pedicle screw-plate system in lower cervical spine,in order to provide basic data for clinical application.

METHODSTotal thirty-two units (functional spinal unit, FSU) were got randomly from 16 cervical speciments, 8 units in each group of C3,4, C4,5, C5,6 and C6,7. The anterior cervical pedicle screw-plate system was implanted to reconstruct the stability of FSU after discectomy and bone graft. The adaptability was measured between the screw-plate system and vertebral body. X-ray and CT were used to evaluate the accuracy of anterior cervical pedicle screws. The subject will be dissected to identify the situation of involvement if screw perforating the pedicle.

RESULTSSixty-four anterior pedicle screws were inserted smoothly in the 32 units. The screw and the plate were harmonious locked in the system. The position and length of all screws were satisfactory through X-ray views. However,6 screws perforated the transpedicular (degree 1) according to CT axial views,2 internally cortex and 4 laterally cortex. None perforation was degree 2 or more. None cervical sac compression and nerve root injury was observed in two internal perforation cadavers. One vertebral vein involvement was found in the four lateral perforation screws. The vertebral artery was not pinched though one screw near to the artery.

CONCLUSIONThe anterior cervical pedicle screw-plate system is adapted to reconstruct in lower cervical spine and it deserved to be used for clinical application.

Aged ; Bone Plates ; Bone Screws ; Cervical Vertebrae ; surgery ; Equipment Design ; Feasibility Studies ; Female ; Humans ; Male ; Materials Testing ; Middle Aged

OBJECTIVETo explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo.

METHODSMelanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

RESULTSThe wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

CONCLUSIONMelanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.

Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Coculture Techniques ; Collagen Type I ; Humans ; Melanocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Nude ; Skin ; injuries ; Skin, Artificial ; Tissue Engineering

OBJECTIVE: To detect porcine endogenous retrovirus (PERV) in Daweizi pigs and to provide basic parameters of evaluating the biological safety for xenotransplantation from pigs to humans. METHODS: Ear tissues from 42 individuals were randomly collected from a Daweizi pig population. PCR and RT-PCR were performed to detect PERV proviral DNA and mRNA respectively. Finally, env-A, env-B, and env-C were amplified, sequenced, and analyzed using the BLAST software in National Center for Biotechnology Information. RESULTS: PERV proviral DNA and mRNA could be detected in the 42 individuals by PCR and RT-PCR, respectively. env-A, env-B and env-C were detected in all the individuals. Compared with other pig species (AY288779, DQ011794 and AY534304), there was 1 and 8 bp differences in the sequences of env-A and env-C, while no difference in env-B. CONCLUSION PERV exists and has transcriptive activity in Daweizi pigs. The predominate subtype is PERV-ABC. Env genes are firstly cloned and sequenced in Daweizi pigs and there are polymorphism in the breed. As to the biological safety, the breed was not suitable as a donor in xenotransplantation.
Animals ; DNA, Viral ; isolation & purification ; Endogenous Retroviruses ; isolation & purification ; Humans ; Polymerase Chain Reaction ; methods ; Swine ; virology ; Transplantation, Heterologous