In order to obtain higher L-lactic acid yield industrial strain, the original strain Rhizopus oryzae PW352 was mutated by means of N+ ions implantation and a mutant strain Rhizopus oryzae RE3303 was obtained. Its lactic acid yield was increased by 75% than that of the original one. The acid producing condition was optimized by orthogonal design. The concentration of L-lactic acid reached to 131～136g/L and the conversion rate of glucose was as high as 86%～90% under the optimum condition.

Objective To investigate the effect of insulin on D_5 dopamine receptor expression and function in renal proximal tubule (RPT).Methods Immortalized RPT cells and D_5 receptor transfected HEK293 (HEK-D_5) cells were used in the study to investigate the effect of insulin on D_5 receptor expression and function,and those effects were compared in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). The function of D_5 receptor was determined by measurement of the Na~+-K~+-ATPase activity in HEK-D_5 cells. Results Insulin increased D_5 receptor protein expression in a concentration and time-dependent manner in WKY RPT cells,but not in SHR.The basal level of D_5 receptor expression was higher in WKY cells than that in SHR cells. Stimulation with fenoldopam(D_1-like dopamine receptor agonist) inhibited the Na~+-K~+-ATPase activity;pretreat- ment with insulin increased the inhibitory effect of fenoldopam on Na~+-K~+-ATPase activity in HEK-D_5 cells. Conclusion The abnormal regulation of insulin on D_5 receptor expression and function might be involved in the path- ogenesis of essential hypertension.

Objective To determine whether the poking reduction and bone grafting technique with guide through bony tunnel can correct a Hill-Sachs lesion. Methods A total of 30 cadaveric humeri were equally divided into three groups, 10 cadaveric humeri per group. Hill-Sachs lesions were replicated with a osseous defect involving 10% (group A ) , 20% (group B ) and 30% (group C ) of the articular surface. All the bone defects in each group were measured and the poking reduction and bone grafting technique with guide through a bony tunnel was performed in group B and group C. The preoperative and postoperative transverse arc length, longitudinal are length, depth and volume of the osseous defects in group B and group C were compared by using paired t test. Results Before reduction, the transverse arc length of the bone defects was ( 10.9 ± 1.4 )mm in group B and ( 16.3 ± 2.3 ) mm in group C ; longitudinal arc length was ( 22.4 ± 2.4 ) mm in group B and ( 28.0 ± 2.2 ) mm in group C ;depth was (6.9±0.9) mm in group B and (11. 1 ±0.9) mm in group C; volume was (708.7±93.9) mm3 in group B and (1338.3 ± 185.6) mm3 in group C. After reduction, the transverse arc length of the bone defects was (5.1 ± 2.4 ) mm in group B and ( 7.6 ± 3.6 ) mm in group C ; longitudinal arc lengthwas (10.5 ±4.9) mm in group B and (12.3 ±5.3) mm in group C; depth was (0.3±0.1 ) mm in group B and (0.4 ±0.1 ) mm in group C; volume was (48.9 ± 16.1 )mm3 in group B and (70.3 ± 37.9) mm3 in group C. The comparison of all the parameters showed statistical difference (P <0. 01 ). Conclusion The poking reduction and bone grafting technique with guide through a bony tunnel can effectively correct the Hill-Sachs lesions with humeral head osseous defects involving 20% -30% of the articular surface.

OBJECTIVETo study directional differentiation of BMSCs guided by Desert living Cistanche (Herba Cistanches) which invigorates the kidney.

METHODSPrimary BMSCs were obtained by whole bone marrow culture and subcultured to the fourth generation by trypsin digestion, and than inoculated into two six-well plates at 5 x 10(6) cells per milliliter, all the plates were divided into three groups as blank group, Dexamethasone (DXM) group and Herba Cistanches group, three wells in each group, medium were changed at day 2. The blank group were changed with L-DMEM containing 10% FBS. The DXM group were changed with medium containing 10 mmol/L beta-sodium glycerophosphate, 0.1 micromol/L DXM and 50 mg/L vitamin C. The Herba Cistanches group were changed with medium containing 10% blood serum containing Herba Cistanches and L-DMEM. One of the six-well plates was stained by alkaline phosphatase (AKP) at the tenth day,the other one was stained by alizarin Bordeaux at the twentieth day.

RESULTSAt the tenth day DXM group and Herba Cistanches group were ALKP stained positive; from the 12th day,white calcium nodus could be seen at the surface of the wells; which alizarin stained positive by the twentyth day.

CONCLUSIONThe medium containing Herba Cistanches can guide BMSCs to differentiate into osteoblast, which promises a favorable prospect for the treatment of osteoporosis and bone fracture disunion.

Animals ; Cell Count ; Cell Differentiation ; drug effects ; Cells, Cultured ; Cistanche ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Rats ; Rats, Sprague-Dawley

Objective To explore the method of repairing wound defects with exposed tibia. Methods 32 patients with soft tissue defects with exposed tibia were treated with three kinds of flaps pedicled with interior,lateral and posterior vascular perforating branches in the leg,respectively.Re- suits One flap with distal part necrotic was treated with change of dressings and got one-stage healing. One case had delayed union for muscular infection under the flap.Other flaps all were successful and sur- vived.No ostemyelitis was found.Conclusion The flap pedicled with vascular perforating branches of leg has abundant blood supply and is a good method for repairing small or middle wound defect with ex- posed tibia.

Adult ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Shoulder Dislocation ; therapy

BACKGROUNDIn China the ginseng root began to be used in medicine over 2000 years ago. Ginsenosides are the most important component isolated from ginseng. The authors investigated the effect of ginsenoside Rg1 on the spectrum of gene expression in the endothelial cells stimulated by TNF-alpha and further explored the potential molecular mechanism of endothelial protection by ginsenoside Rg1.

METHODSNitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by using an NO assay kit. A home-made oligonucleotide microarray containing approximately 400 cardiovascular disease-related genes was constructed. The alteration of the spectrum of gene expression induced by ginsenoside Rg1 in HUVECs which were activated by TNF-alpha were detected by oligonucleotide microarray analysis.

RESULTSNO production in HUVECs was decreased significantly after TNF-alpha treatment, while pretreatment with ginsenoside Rg1 enhanced NO production in TNF-alphastimulated HUVECs. Ginsenoside Rg1 affected the expression levels of genes involved in vascular constriction, cell adherence, coagulation, cell growth and signal transduction in TNF-alphastimulated HUVECs.

CONCLUSIONSGinsenoside Rg1 could enhance NO production and the expression of eNOS mRNA in TNF-alpha stimulated HUVECs. Ginsenoside Rg1 regulated sets of genes in endothelial cells and protected endothelial cells from TNF-alpha activation. Microarray analysis provided us with valuable insights into the atheroprotective mechanism by gingsenoside Rg1.

Endothelial Cells ; drug effects ; physiology ; Gene Expression ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; analysis ; Nitric Oxide Synthase Type III ; Oligonucleotide Array Sequence Analysis ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the effect of gene expression of mouse uroplakin II (UPII) promoter on human bladder cell cancer cell line.

METHODSThe mRNA expression of different cell lines was quantified by RT-PCR. Green fluorescent protein (GFP) and luciferase (Luc) were used as reporter genes. The plasmids carrying UPII or GFP were constructed and transfected into human cell lines of bladder transitional cell cancer (BIU-87), kindey cancer (GRC-1), vascular endothelium (EC), lung cancer cell line (A549) and skin fibroblast cell line (Hs27). GFP activity of cells was detected by confocual microscopy and flow cytometry (FCM). Luciferase value was measured by luminometer (microplate) and luciferase to beta-galactosidase ratios (L/G values) were used for evaluating transfection efficiency.

RESULTSRT-PCR showed high expression level of UPII mRNA in bladder cancer cell line BIU-87, whereas low level or no expression in nonbladder cancer cell lines. The activity of GFP in bladder cancer (BIU-87) cell was higher than that in the other cell lines (5 - 10/HP versus 0 - 2/HP), with 4.34% positive cells in BIU-87 detected by FCM, but no positive cell was found in the other cell lines. L/G values indicated that the luciferase expression in human bladder cancer cells transfected with mouse UPII promoter was 1.8 - 8.2-fold as high as that in the nonbladder cell lines.

CONCLUSIONMouse UPII promoter gene can be expressed in a tissue-specific fashion in human urinary bladder cancer. It is capable of initiating transcription of reporter genes in human bladder cancer cell line.

Animals ; Cell Line, Tumor ; Flow Cytometry ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Membrane Proteins ; genetics ; Mice ; Organ Specificity ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Urinary Bladder Neoplasms ; genetics ; therapy ; Uroplakin II

OBJECTIVETo explore the role of inflammatory reaction mediated by p38-mitogen activated protein kinase (p38-MAPK) signal path on prevention and treatment of Parkinson disease (PD) model rats by electroacupuncture (EA).

METHODSThirty-two healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, eight rats in each one. The PD model was established in the model group and EA group by subcutaneous injection of rotenone in skin-back area (2 mg/kg, dissolved in sunflower oil, 2 mg/mL in density), while the injection of sunflower oil emulsion without rotenone at the same point and quantity as the model group was applied in the sham operation group. The normal group was not given any intervention. The EA treatment (continuous wave, 2 Hz in frequency, 1 mA in intensity, 20 min) was applied at "Fengfu" (GV 16) and "Taichong" (LR 3) in the EA group, once a day for continuously 14 days. No treatment was given in the other groups. The expression of tyrosine hydroxylase (TH), phosphorylated p38-MAPK, cyclooxygenase-2 (COX-2) in the substantia nigra were detected with immunohistochemical method.

RESULTSThere was typical PD ethology change in the model group. Compared with the normal group and sham operation group, the expression of TH positive neuron in the substantia nigra in the model group was significantly decreased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly increased (all P < 0.01). Compared with the model group, the expression of TH positive neuron in the EA group was apparently increased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly decreased (all P < 0.01).

CONCLUSIONThe EA therapy could obviously reduce the expression of inflammation mediator COX-2, inhibit the phosphorylation of p38-MAPK, reduce the damage of dopaminergic neurons in the rats with PD, and this effect may be related with the impact of p38-MAPK signal path

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Electroacupuncture ; Humans ; Male ; Parkinson Disease ; enzymology ; genetics ; therapy ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; enzymology ; Tyrosine 3-Monooxygenase ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo develop the characteristic chromatographic profile of Sarcandra glabra by HPLC for its quality control.

METHODThe HPLC analysis was performed on an Agilent Zorbax Eclipse XDB-C18 column (4. 6 mm x 250 mm, 5 microm) with column temperature at 40 degree C. The mobile phase was consisted of water containing 0. 5% formic acid and acetonitrile to methanol (1:9) in gradient mode, and the detection wavelength was set at 344 nm.

RESULTA common mode of the HPLC characteristic chromatographic profile has been establised. There were 20 common peaks , seven of which were identified, and 46 samples from different habitats were classified into five groups based on principal component cluster analysis.

CONCLUSIONThe method was time-saving and can represent the chemical information and provide a scientific basis for quality control of S. glabra.

Chromatography, High Pressure Liquid ; Cluster Analysis ; Drugs, Chinese Herbal ; chemistry ; standards ; Ferns ; chemistry ; Organic Chemicals ; analysis ; chemistry ; isolation & purification ; Quality Control