A strain of yeast capable of hydrolyzing ethyl ester of racemic Ketoprofen with high enantioselectivity has been isolated from soil after two-step enrichment. The yeast was identified as Trichosporon brassicae. The process of growth and enzyme production was investigated. The catalytic performance of the resting cell of KET4 in kinetic resolution of Ketoprofen was also investigated. When the conversion of substrate reached 41% , enantiomeric excess of the (S) - Ketoprofen produced was 91 % , indicating a high enantiomeric ratio of 45.

OBJECTIVETo investigate the effect of noninvasive positive pressure ventilation( NIPPy) on the gene and protein expression of biquitin-proteasome of skeletal muscle in patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD).

METHODSSeven patients with AECOPD by NIPPV were used as the study group, meanwhile, 6 patients with AECOPD who refused NIPPV was the control group. The blood gas analysis, heart rate (HR) and mean arterial pressure (MBp) were monitored before and 14 days after treatment. A skeletal muscle biopsy was performed after 14 days of therapy. The mRNA expression of ribosomal protein S21 (RPS21), Ubiquitin, Ubiquitin combined with enzyme E2 (E2), Ubiquitin ligase E3 (E3) in skeletal muscle cell were measured by RT-PCR. The protein expression of mitochondrial aconitase (AC02), protease C3 (C3), ribosomal protein SLC16 (SLC16) were detected by Western blot.

RESULTSForteen days after treatment, the patients in NIPPV group got much better improvement in PaCO2, PaO2 and HR than that of the patients.in the control group (P < 0.05). The gene expression of RPS21,Ubiquitin, E2 and E3 in skeletal muscle cell on patients with NIPPV were obviously lower than that of the control group (P < 0.05, P < 0.01). Compared with that of the control group, the protein expression of C3 and AC2 increased significantly in the NIPPV group (P < 0.01). The protein expression of SLC16 was significantly lowered in the treated group (P < 0.01).

CONCLUSIONNIPPV can ameliorate the proteasome pathway and energy metabolic disorders in patients with AECOPD.

Humans ; Muscle, Skeletal ; metabolism ; Positive-Pressure Respiration ; Proteasome Endopeptidase Complex ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; therapy ; Ubiquitin ; metabolism

OBJECTIVETo observe the effect of natural type ginsenoside Rg2 (Rg2) and its stereoisomers [20 (R)-Rg2 and 20 (S)-Rg2] at different concentrations on oxygen-glucose deprivation/ reperfusion (OGD/R) induced cortical neuronal injury model in vitro, and to explore the mechanism, and compare their differences of action.

METHODSCortical neurons after 7-day culture were randomly divided into 5 groups, i.e., the control group, the model group, the Rg2 group, 20 (R) -Rg2 group, and 20 (S) - Rg2 group. Cortical neurons in the Rg2 group, 20 (R)-Rg2 group, and 20(S)-Rg2 group were pretreated with 20, 40, and 80 μmol/L Rg2, 20 (R) -Rg2, and 20 (S) -Rg2 for 24 h to prepare OGD/R model. The cell survival rate, the activity of Caspase-3, the intracellular Ca2+ concentration, contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected 24 h later.

RESULTSCompared with the control group, cell survival rates and activities of SOD obviously decreased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly increased with statistical difference (P < 0.05). Compared with the model group, cell survival rates and activities of SOD obviously increased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly decreased in 20 μmol/L Rg2 group, 40 μmol/L 20 (R) -Rg2 group, and 80 μmol/L 20 (S) -Rg2 group (P < 0.05). Compared with 20(S)-Rg2 group, cell survival rates increased and contents of MDA significantly decreased in 20, 40, and 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). The activity of Caspase-3 decreased and contents of SOD increased in 80 μmol/L 20 (R)-Rg2 group, and 40, 80 μmol/L Rg2 groups (P < 0.05). Ca2+ fluorescent optical gray value decreased in 40, 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). Compared with 20 (R)-Rg2 group, Ca2+ fluorescent optical gray value decreased in 80 μmol/L Rg2 group (P < 0.05); contents of SOD increased in 40 and 80 μmol/L Rg2 groups (P < 0.05); contents of MDA decreased in 20, 40, and 80 μmol/L Rg2 groups (P < 0.05).

CONCLUSIONSRg2 and its stereoisomers could improve cell vitality of cortical neurons against OGD/R induced injury. This might be related to improving anti-apoptotic capacities and antioxidant abilities, and reducing Ca2+ inflow. Besides, the neuroprotective effect of 20 (R) -Rg2 was better than that of 20 (S) -Rg2, but inferior to that of Rg2.

Antioxidants ; metabolism ; Apoptosis ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; Cells, Cultured ; Ginsenosides ; pharmacology ; Glucose ; Humans ; Malondialdehyde ; metabolism ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Oxygen ; Random Allocation ; Reperfusion Injury ; Stereoisomerism ; Superoxide Dismutase ; metabolism

Objective To discuss the value and experience of the percutaneous vertebroplasty (PVP)in the treatment of vertebral body compression fracture(VCF)in aged osteoperosis.Methods PVP was performed in 44 cases with VCF including 28 with single vertebral compressed fracture,12 with double compressed fracture and four with triple compressed fracture,with 67 vertebrae,for clinical and radiologieal evaluation.Results The mean follow-up was 15 months(4-23 months).There could be seen immediate relief of pain in 40 cases,out-of-bed activities at operation day in 19 and out-of-bed activ- ities at second day after operation in 25.Postoperative X-ray showed uniformly distributed bone cement in the vertebral,without leakage.Conclusion PVP is a recommendable method for VCF,for it has ad- vantages of pain relief,vertebrae stabilization,minimal invasion and minor complications.

Objective To establish a convenient,accurate and practical method for detection of adefovir dipivoxil resistance-as- sociated mutation in hepatitis B virus:rtA181V/T/S and rtN236T mutations.Methods According to HBV complete sequences in GenBank,two pairs of primers were designed to amplify the region of HBV reverse transcriptase in order to introduce a BglI restriction site upon PCR product of wild type (wt) and a BseDI restriction site upon PCR product of rt236 mutant type.After amplification,the PCR products were digested with BglI and BseDI separately.We used this method to detect wild,rt181 mu- tant,rt236 mutant plasmids and 3 chronic hepatitis B patients' serum with obvious ADV resistance-associated mutations.We also tested the sensitivity of this method by mixing the wild and mutant plasmids in different proportions.Results The method could detect rt181 and rt236 mutations simultaneously.The result of RFLP analysis was in accordance with that of DNA se- quencing and cloning analysis.This method could detect the mutants even when they comprised only 10% of the total virus population.Conclusions The PCR-RFLP method with high sensitivity can detect rt181 and rt236 mutations simultaneously.It can be used for early detection of ADV resistance-associated mutation in hepatitis B virus.

BACKGROUNDPreparing hyperthyroid patients for thyroid surgery with a combination of antithyroid drugs and thyroxine has long been controversial because this combination usually results in only partial inhibition of thyroid function. We therefore used large doses of antithyroid drugs to completely inhibit the synthesis of thyroxine and render the thyroid gland defunctionalized. We then administered physiologic doses of thyroxine to inhibit thyroid-stimulating hormone secretion. We have named this treatment "sequential thyroid defunctionalization followed by thyroxine supplementation."

METHODSFour hundred and seventy-one hyperthyroid patients seen at our hospital were divided into experimental and control groups. The control group was treated preoperatively with antithyroid drugs and iodine preparation. The experimental group was further divided into four subgroups and treated with "sequential thyroid defunctionalization followed by thyroxine supplementation". Each of the four subgroups received different doses of antithyroid drugs and thyroxine for differing time periods. Thyroid function was assessed at each stage of treatment, as were operative blood loss volumes and postoperative complications.

RESULTSCompared to the control group, the four experimental groups showed less thyroid congestion and surface varices at surgery. Patients in subgroup A also had thyroid glands that were almost histologically normal. The mean operative blood loss volume of the experimental group was less than that of the control group (326 +/- 163) ml in the control group; (196 +/- 57) ml in subgroup A; (230 +/- 71) ml in subgroup B; (240 +/- 80) ml in subgroup C; and (312 +/- 97) ml in subgroup D). The postoperative complication rate of the experimental group was 8.64% (21/243) whereas that of the control group was 17.54% (40/228).

CONCLUSIONSSequential thyroid defunctionalization followed by thyroxine supplementation is effective in reducing the bleeding volume and postoperative complication rate in selected hyperthyroid patients undergoing thyroidectomy.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Hyperthyroidism ; physiopathology ; surgery ; Male ; Middle Aged ; Thyroid Gland ; pathology ; physiopathology ; Thyroidectomy ; Thyroxine ; administration & dosage

OBJECTIVETo design a clinically applicable transoralpharyngeal atlantoaxial reduction plate (TARP), introduce the operation procedure, and evaluate its preliminary clinical effects.

METHODSA novel TARP system, including butterfly titanium alloy plate, self-locking screws, atlantoaxial reductor and other operational instruments was developed. This system was applied clinically on five patients with irreducible atlantoaxial dislocation of congenital or traumatic origin. During operation, the reduction was completed by the combined action of the plate and the atlantoaxial reductor after transoral joint release and cord decompression. Bone graft granules were implanted between the bilateral atlantoaxial joints and TARP was used to immobilize subsequently the atlas and axis.

RESULTSClinical application demonstrated that TARP could induce instant reduction and that the method was operationally feasible and its postoperational effect was satisfactory.

CONCLUSIONSThe design of TARP is novel. The operational procedure is simple and easy to use. Furthermore, instant reduction can be completed during the operation and the fixation is relatively stable. TARP is an ideal alternative for irreducible atlantoaxial dislocation and may have excellent prospects for further clinical applications.

Adolescent ; Adult ; Atlanto-Axial Joint ; surgery ; Bone Plates ; Bone Screws ; Decompression, Surgical ; methods ; Equipment Design ; Female ; Humans ; Internal Fixators ; Joint Dislocations ; surgery ; Male ; Mouth ; surgery ; Spinal Fusion ; methods

OBJECTIVETo investigate the changes of thyroxin and monocyte human leukocyte antigen-DR expression in senior patients with sepsis and explore their clinical significance.

METHODSAccording to the 2001 SCCM/ESICM/ACCP/ATS/SIS international sepsis definitions, 125 senior patients with sepsis free of thyroid conditions were divided into non-severe sepsis group (n=86) and severe sepsis group (n=39), with another 30 healthy subjects as the control. Thyroid function was assayed by chemoluminescence method in these patients and monocyte HLA-DR expression was determined by flow cytometry.

RESULTSCompared with the control group and non-severe sepsis cases, the levels of free T3 (FT3), free T4 (FT4), T3, T4 and monocyte HLA-DR expression were significantly lower in severe sepsis cases (P<0.05), but the levels of thyroid stimulating hormone (TSH) were comparable between the 3 groups (P>0.05). The non-severe sepsis cases showed significantly lower levels of FT3, FT4, T3, T4, TSH and monocyte HLA-DR expression than the control group (P<0.05). In severe sepsis group, the levels of FT3, FT4, T3, T4 and monocyte HLA-DR expression showed significant differences between the fatal cases and surviving cases (P<0.05).

CONCLUSIONThe levels of thyroxin and monocyte human leukocyte antigen-DR expression are obviously lower in senior patients with severe sepsis, and their detection may well indicate the severity of the condition and help make prognostic judgment.

Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; HLA-DR Antigens ; blood ; immunology ; Humans ; Male ; Monocytes ; metabolism ; Pneumonia ; complications ; Sepsis ; blood ; etiology ; immunology ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo study the resistant rate of hepatitis B virus (HBV) to ADV and the dynamic evolution of HBV in lamivudine (Lam)-resistant chronic hepatitis B (CHB) patients.

METHODSTwenty-three Lam-resistant CHB patients were assigned to a 10mg/d ADV monotherapy for 68-116 weeks. The baseline and different time point blood samples after ADV monotherapy were analyzed for ADV-resistant mutations using direct sequencing of PCR products; the evolution of HBV mutations was examined by clonal analysis of serial samples from one patient infected with ADV-associated resistant HBV strains.

RESULTSThe cumulative incidence of genotypic ADV resistance at weeks 48 and 96 was 4.3% and 10.5% respectively respectively. The evolution analysis of HBV mutant strains in an ADV-resistant CHB patient showed that the proportion of YMDD mutants gradually decreased with rtA181S mutants increasing over time after ADV monotherapy, and that rtA181S+N236T mutants became the predominant strains during prolonged ADV monotherapy. The addition of Lam to the ongoing ADV treatment had poorer antiviral response in the patient with rtA181S or rtA181S+N236T mutant infection; one clone with multi-drug resistant mutations was selected during Lam and ADV combination therapy.

CONCLUSIONIncreased risk of adefovir resistance and selection of multi-drug resistant mutations are associated with long-term ADV monotherapy in patients with Lam-resistant chronic hepatitis B.

Adenine ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; therapeutic use ; Drug Resistance, Viral ; Evolution, Molecular ; Female ; Hepatitis B virus ; classification ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; Male ; Middle Aged ; Organophosphonates ; therapeutic use

With the development of molecular biology and genomics,metagenomics is playing a more important role in forensic science and forensic identification.In recent years,as a branch discipline studying the composition profile and diversity of microbe flora as well as studying the interaction within microbe and with environment,the application of metagenomics has gradually risen and brought new opportunities for forensic identification-related area.In this review,strategy of metagenomics and its application in forensic identification including individual identification,origin determination of biological stain in crime scene and drug abuse detection are summarized.This article aims to elucidate the role and application value of metagenomics in forensic science.