The 1,095 bp gene encoding peroxidase from Coprinus cinereus was synthesized and integrated into the genome of Pichia pastoris with a highly inducible alcohol oxidase. The recombinant CiP (rCiP) fused with the a-mating factor per-pro leader sequence derived from Saccharomyces cerevisiae was secreted into the culture medium and identified as the target protein by mass spectrometry, confirming that a C. cinereus peroxidase (CiP) was successfully expressed in P. pastoris. The endoplasmic reticulum oxidoreductase 1 (Ero1) and protein disulfide isomerase (PDI) were co-expressed with rCiP separately and simultaneously. Compared with the wild type, overexpression of PDI and Erol-PDI increaseed Cip activity in 2.43 and 2.6 fold and their activity reached 316 U/mL and 340 U/mL respectively. The strains co-expressed with Erol-PDI was used to high density fermentation, and their activity reached 3,379 U/mL, which was higher than previously reported of 1,200 U/mL.
Coprinus ; enzymology ; Culture Media ; Cytoplasm ; Fermentation ; Glycoproteins ; metabolism ; Mass Spectrometry ; Mating Factor ; Oxidoreductases Acting on Sulfur Group Donors ; metabolism ; Peptides ; Peroxidases ; biosynthesis ; Pichia ; metabolism ; Protein Disulfide-Isomerases ; metabolism ; Protein Folding ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae Proteins ; metabolism

Objective To study the epidemiological and clinical characteristics of patients with severe acute respiratory syndrome(SARA). Methods Clinical data from 682 patients with SARS were retrospectively analyzed. Results The patients ranged from 13 to 76 years old, 387 male and 295 female. A total of 356 patients (52.2) had a history of close contact with diagnosed SARS patients, and 113 (25.1%) had been to hospital ever. The most common symptom was fever (99.3%), followed by cough (44.4%), shortness of breath (12.2% ), diarrhea (8.9%). 596 patients (87.4%) had normal or decreased white blood cell counts. Serum ALT and CPK levels were elevated in 112 patients (16.4%)and 17 patients (2.5%) respectively. Infiltrates on chest radiography were seen in all patients, with 69.8% involved both lungs. Six patients (0.9%) died of SARS. Conclusion SARS is infectious. Hospital is an important place where SARS transmits. Fever, cough and infiltrative changes on chest radiography are mainly symptoms and signs.

Background: Dampness-heat syndrome is a major syndrome type in patients with a sharp deterioration of chronic renal failure (CRF). Qingshen Decoction, a compound traditional Chinese herbal medicine, could relieve the clinical symptoms of CRF patients, and was considered to have a certain reversal effect on rapid deterioration of renal function. Objective: To observe the changes of serum interleukin-8 (IL-8) and IL-18 levels in CRF patients with a sharp deterioration and dampness-heat syndrome, and to explore the curative efficacy of Qingshen Granule. Design, setting, participants and interventions: Sixty CRF patients with a sharp deterioration and dampness-heat syndrome from Department of Nephrology, the First Affiliated Hospital of Anhui College of Traditional Chinese Medicine, were randomly divided into treatment group (30 cases) and control group (30 cases), with another 20 healthy individuals as normal control. The patients in the treatment and control groups were all treated with Jiedu Xiezhuo II, a compound traditional Chinese herbal medicine, given as retention enema. Qingshen Granule was additionally administered to the patients in the treatment group with 1 dosage each time and 3 times a day. The treatment course was one month. Main outcome measures: The levels of serum IL-8 and IL-18 in the normal individuals and before and after treatment in the treatment and control groups were detected. Results: The total response rates of treatment group in clinical efficacy assessment and assessment of syndrome of traditional Chinese medicine (86.67% and 86.67%) were higher than those of the control group (56.67% and 60%), and there were significant differences between the two groups (P<0.05). The levels of serum IL-8 and IL-18 in CRF patients before treatment were obviously higher than those in the healthy individuals (P<0.01), and there were no differences in the levels of IL-8 and IL-18 before treatment between the treatment group and control group (P>0.05). After one-month treatment, the levels of serum IL-8 and IL-18 were markedly decreased in the two groups (P<0.01), and the levels of serum IL-8 and IL-18 in the treatment group were markedly lower than those in the control group (P<0.05). There were differences in decreased degrees of IL-8 and IL-18 levels between the two groups (P<0.05). Conclusion: CRF patients with a sharp deterioration and dampness-heat syndrome have high serum IL-8 and IL-18 levels. Qingshen Granule can reduce the levels of serum IL-8 and IL-18, and improve the renal function and ameliorate the clinical symptoms.

OBJECTIVETo evaluate biomechanical properties in different methods of internal fixation combined with distal tibiofibular syndesmosis injury, in order to provide a theoretical basis for clinical choice.

METHODSSix lower limbs specimens were collected and divided into 5 groups, including normal group, distal tibiofibular syndesmosis injury (injury group), 3 cortexes group, 4 cortexes group and hook-plate fixation group. Neutral position, plantar flexion position (30°), dorsiflexion (20°) supination external rotation position of foot movement were simulated on universal materials tester. Strength, stiffness and stability of ankle joint in 4 kinds of motion conditions were measured.

RESULTSThere was significant differences in strength and stiffness of ankle joint between injury group and normal group in 4 different kinds of motion conditions (P<0.05). Strength and stiffness of ankle joint in 3 cortexes group, 4 cortexes group and hook-plate fixation group were improved obviously in 4 different kinds of motion conditions, and biomechanical indexes were recovered normally or better than normal group. Stiffness of the three fixation groups were better than normal group,but there was no significant differences among three groups (P > 0.05), while stiffness of hook-plate fixation group was closed to normal group.

CONCLUSIONFor distal tibiofibular syndesmosis injury, 1 screw with 3 cortexes, 4 cortexes and hook-plate had a positive impact on strength, stiffness and stability of distal tibiofibular syndesmosis injury stress,and could restore the normal stabllity of ankle joint after reconstruction. While fixing by screw fixation would limit the rototary motion of ankle joint,ankle mortise could not adapt to changes of talus bone, thus induces screw breakage and traumatic arthritis. Hook-plate fixation is more suitable than 3 cortexes or 4 cortexes fixation for bilmechanical properties,and its' stress is more balance and can reduce postoperative complcations.

Ankle Injuries ; physiopathology ; surgery ; Biomechanical Phenomena ; Fibula ; injuries ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Joint Instability ; physiopathology ; prevention & control ; Reconstructive Surgical Procedures ; methods ; Tibia ; injuries ; surgery

Objective To investigate the hemodynamics of increasing portal venous pressure(PVP) in hepatocellular carcinoma patients complicated with hepatic arterioportal vein fistulas (HAPVF)and the diagnosis and therapy of intractable upper gastrointestinal hemorrhage caused by HAPVF.Methods One hundred and fifteen cases of hepatocellular carcinoma with upper gastrointestinal hemorrhage were checked by hepatic arteriography and were treated through orifices embolization in cases with severe HAPCF by coils and/or ethanol. Results Twenty-six out of 31 patients suffering intractable upper gastrointestinal hemorrhage have severe HAPVF(the main stem of portal veins are visible).However,there are only 15 patients with light HAPVF among the 84 patients who have mild upper gastrointestinal hemorrhage (the main stem of portal veins are invisible).After the embolization,all of the 26 patients who have severe HAPVF stopped bleeding.Among them,the main stem of hepatic arteries are occluded in 2 patients. Conclusion The existence of severe HAPVF should be taken into consideration when intractable upper gastrointestinal hemorrhage occurs in hepatocellular carcinoma patients,and it can be diagnosed through hepatic artery DSA.Orifice embolization is the most effective method for such kind of hemorrhage.

To evaluate the efficacy of Tuina and Chinese patent drug Shuxuetong injection in preventing patients undergoing total knee arthroplasty from deep venous thrombosis and in functional rehabilitation.

AIM: To investigate the effect of curcumin on oxidized low-density lipoprotein (ox-LDL)-induced injury of human aortic endothelial cells (HAECs).METHODS: HAECs were pre-treated with curcumin at different concentrations and then treated with ox-LDL.The cell viability was assessed by MTT assay.The cell proliferation ability was analyzed by EdU assay.ELISA was used to determine the concentrations of interleukin-6 (IL-6), transforming growth factor β1 (TGFβ1), high mobility group box-1 protein (HMGB1) and secretory receptor for advanced glycation end products (sRAGE) in the HAEC culture medium.The binding activity of peroxisome proliferator-activated receptor γ (PPARγ) was evaluated by electrophoretic mobility shift assay.The protein levels of HO-1, HMGB1, RAGE,IL-6,TGFβ1 and phosphorylated PPARγ in the HAECs were determined by Western blot.RESULTS: The viability and the proliferation ability decreased significantly in the HAECs treated with ox-LDL.The PPARγ/HO-1 signaling pathway was inhibited while its down-stream HMGB1/RAGE signaling pathway was activated by ox-LDL.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were increased.Pre-treatment with curcumin activated PPARγ/HO-1 signaling pathway and inhibited HMGB1/RAGE signaling pathway in ox-LDL treated HAECs in a concentration-dependent manner.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were also decreased dramatically by pre-treatment of curcumin in a concentration-dependent manner.CONCLUSION: ox-LDL induces HAEC damage by inhibiting PPARγ/HO-1 to activate HMGB1/RAGE inflammatory signaling.Curcumin exerts protective effect on ox-LDL treated HAECs via activating PPARγ/HO-1 signaling pathway.

OBJECTIVETo investigate the antibacterial activity of decoction of Radix glycyrrhizae against Streptococcus mutans (S. mutans) in vitro.

METHODSThe decoction of Radix glycyrrhizae was prepared by boiling particles of Radix glycyrrhizae, the diameter was 0.2-3.2 mm. In distilled water and filtered, the filtrate was collected for study. The minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) of the decoction against S. mutans were detected using double dilution. The effect of decoction on growth and acidogenic profile of S. mutans were investigated by detecting the Abs of bacteria suspension and the pH value of medium at definite time intervals(0, 3, 7, 12, 23, 40 h) during cultured.

RESULTSThe MIC determined for decoction was 50 mg x mL(-1) and there was no bactericidal effect when concentration of decoction lower than 100 mg x mL(-1). The decoction inhibitted multiplication of bacteria significantly and the effects became stronger with concentration increasing. The decoction also inhibitted S. mutans producing acid and the effect became stronger with concentration increasing. The most efficient inhibition were observed when incubated 12 hours.

CONCLUSIONThe decoction of Radix glycyrrhizae can inhibite the growth and acid-production of S. mutans in vitro.

Anti-Bacterial Agents ; Bacteria ; In Vitro Techniques ; Microbial Sensitivity Tests ; Plant Extracts ; Streptococcus mutans

OBJECTIVETo establish an efficient and stable method for protein extraction of Streptococcus mutans.

METHODSThe collected bacteria were treated by freeze-thaw and ultrasonic (method 1), ultrasonic (method 2), boiling (method 3), boiling and ultrasonic (method 4), respectively. The index such as state of bacteria broken, concentration of extracted protein and SDS-PAGE of protein were employed to evaluate the effects of above four methods.

RESULTSBeside the method 3, the other three methods could break the bacteria effectively, of which ultrasonic was the key factor. The pattern of SDS-PAGE which treated by method 1, method 2 and method 4 was almost same, but method 1 resulted in the best abundance. There was significantly difference among the four protein concentration extracted by four methods (P < 0.05). All methods exhibited good stability and reproducibility.

CONCLUSIONMethod of freeze-thaw and ultrasonic resulted in an efficient proteins extraction of Streptococcus mutans which demonstrated good stability and reproducibility and easy to handle.

Bacterial Proteins ; Reproducibility of Results ; Streptococcus mutans

OBJECTIVETo explore the operative methods and effects of treatment of old acetabular fractures.

METHODSFrom October 2001 to October 2007, 26 patients with old acetabular fractures were treated with operation including 21 males and 5 females with an average age of 34 years ranging from 18 to 65 years. On the basis of the three-dimensional computed tomography, all cases were diagnosed and classified according to Letourne-Judet classification, 9 cases were posterior wall fracture, 3 cases were lateropulsition fracture, 7 cases were lateropulsition and posterior wall fracture, 2 cases were posterior column and posterior wall fracture, 2 were T-shape fracture, 3 were dual column fracture. These patients were treated through the anterior,posterior, combined anterior-posterior approaches. The time from injured to operation was 33 to 141 days (averaged 36.4 days). All the fractures were fixed with screws and AO reconstruction plates.

RESULTSAll patients were followed up for 6 to 96 months, with an average time of 32.4 months. Evaluated according to Matta criteria, the results of scores was (5.04 +/- 1.04) on pain, (5.23 +/- 0.76) on range of motion, (4.92 +/- 1.16) on walking,and tatal (5.06 +/- 0.99) on average; The functional results of hip joints were excellent in 6 cases, good in 10 cases, fair in 6 cases, and poor in 4 cases. Sciatic nerve injury was found in 2 patients,lateral femoral cutaneus nerve injury in 3 patients, necrosis of femoral head in 1 patient,infection in 1 patient, and ectopic bone formation in 6 patients.

CONCLUSIONGood clinical results can be obtained by careful selection of operative indications of old acetabular fractures in combination with proper operative approach and correct reduction and fixation.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Time Factors ; Young Adult