The transplants of one-year-old Glycyrrhiza uralensis Fisch. were subjected to five concentrations of MnSO4-H2O (0, 1.81, 18.1, 36.2 and 54.3 mg·L(-1)) culturing in vermiculite. qRT-PCR and HPLC were respectively used to measure the relative expression of SQS1 gene and the content of glycyrrhizic acid of G. uralensis in different concentrations of MnSO4·H2O. This is to explore discuss the effects of the expression of SQS1 gene and the accumulation of glycyrrhizic acid by Mn treatment. The results showed both the expression of SQS1 gene and the content of glycyrrhzic acid of G. uralensis tended to rise after the fall of the first with the increase of concentration of Mn treatment. And they were of very significant positive correlation (P<0.01, r=0.737). Relative expression of SQS1 gene reached the highest 7.90 under 18.1 mg·L(-1) MnSO4·H2O treatment. It was very significantly different between 18.1 mg·L(-1) concentration of MnSO4·H2O treatment and CK (0 mg·L(-1)), 1.81, 36.2 and 54.3 mg·L(-1) (P<0.01), and 1.75, 1.37, 1.37, 2.33 times respectively. The content of glycyrrhizic acid reached the highest under 1.81 and 18.1 mg·L(-1) MnSO4·H2O treatment, and there were not significant difference (P>0.05). It was very significantly different between them and other concentrations of MnSO4·H2O treatment (P<0.01). This study suggests the appropriate concentration of Mn treatment could certain promote the expression of SQS1 gene and the accumulation of glycyrrhizic acid of G. uralensis.
Chromatography, High Pressure Liquid ; Genes, Plant ; Glycyrrhiza uralensis ; chemistry ; genetics ; Glycyrrhizic Acid ; analysis ; Manganese

Animals ; Cardiovascular System ; physiopathology ; Fatigue ; drug therapy ; Mice ; Oxidative Stress ; Periplaneta ; chemistry ; Physical Conditioning, Animal

Objective To analysis and compare the content differences of three effective components of taxanes of Taxus madia Rehd. from five different habitats and growth years by High Performance Liquid Chromatography (HPLC). Methods The mixed liquor of CH3OH and CHCl3 was used for the ultrasonic extraction of 10-deacetyl-bacratin III (10-DAB III), cephalomannine and taxol before being dissolved by CH3OH to obtain sample solution. By HPLC, the contents of three effective components were performed and compared. Results There were extremely significantly differences (P < 0.01) among the content of three effective components of taxanes of Taxus madia Rehd. from different habitats and growth years. Three different habitats and growth years of 10-DAB III, cephalomannine, taxol, and taxanes that have relative higher total content were HK4 > SM3 > SM2, ZL3 > HK4 > ZL4, ZL3 > SM2 > HK4, HK4 > ZL3 > ZL6, respectively. Except for the contents of 10-DAB III and cephalomannine, there were significant positive correlations among the contents of other components (P < 0.01). The contents of taxol would rise or decline when the total contents of three kinds of taxanes rised or declined among different habitats and growth years. The clustering analysis results showed that the contents of taxol have certain peculiarity. In Fuzhou of Fujian, while the total contents of three kinds of taxanes were different from others in Kaifen of Henan. Conclusion Habitants and growth years have a significant impact on the contents change of three effective components of taxanes from Taxus madia Rehd.. Therefore, regulation and control of biosynthetic pathway by choosing habitat conditions and harvest years should be considered, in order to obtain more crude drug of taxol in T. madia.

Objective To explore the clinical application value of multi-slice spiral CT angiography.Methods 28 patients who were suspected cerebrovascular disease were taken cerebrovascular enhanced scan by 16-slice spiral CT,and VR,MIP and MPR post -processing were uset to reconstmct the cerebrovascular image.Results In 28 patients,11 cases were normal,17 cases were abnormal,and among these patients there were 8 cases of abnormal blood vessels,4 cases of aneurysms,1 case of cerebral arteriovenous malformation,4 cases of cerebral artery stenosis.Conclusion The multi-slice CT compared with ordinary spiral CT had the advantage of faster and more secure.It had an important role in guiding the screening and clinical treatment programs in cerebral infarction patients with suspected cerebral artery stenosis.It had a lot of advantages in diagnosis vascular disease and clinical observation after treatment.

OBJECTIVETo study clinical effects of unilateral pedicle screw-rod system fixation combined with the single the cage of interbody fusion in the treatment of lumbar degenerative instability via paraspinal intermuscular approach under MAST Quadrant retractor.

METHODSFrom February 2010 to December 2011, 39 patients with lumbar degenerative instability after invalid conservative treatment were treated with unilateral fixation combined with interbody fusion via the paraspinal intermuscular approach under MAST Quadrant retractor. The indexes such as the operative time, blood loss, complications, VAS pain scores, JOA (subjective symptoms of low back pain,lower limb pain and numbness, sensory disturbance, movement disorders) scores, modified Macnab criteria for curative effect evaluation were observed before treatment, at the 1st month after treatment and the latest follow-up. The intervertebral fusion was studied on the X-ray at the latest follow-up.

RESULTSAll the patients were followed up, and the mean during was (22.3 +/- 8.6) months. The operation time was (138 +/- 46) min, and the amount of bleeding was (335 +/- 152) ml. There were no complications such as cerebrospinal fluid leakage and spinal nerve injury during operation, and no incision infection after operation. The VAS pain score was reduced from preoperative 7.93 +/- 1.27 to 2.05 +/- 1.18 on the 1st month after operation and 1.89 +/- 0.42 at the latest follow-up. The JOA total score was improved from preoperative 1.59 +/- 0.42 to 8.86 +/- 0.37 on the 1st month and 9.02 +/- 0.29 at the latest follow-up. According to modified Macnab criteria, there were 17 cases got an excellent result, 19 good, 3 case fair. Thirty-five patients got intervertebral bony fusion at the latest follow-up.

CONCLUSIONUnilateral pedicle screw-rod system fixation combined with single cage interbody fusion in the treatment of lumbar degenerative instability via paraspinal intermuscular approach under MAST Quadrant retractor is a safe, minimally invasive, satisfactorily effective methods to treat lumbar degenerative instability.

Adult ; Aged ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Intervertebral Disc Degeneration ; surgery ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Reconstructive Surgical Procedures ; Spinal Fusion ; Treatment Outcome

AIM: To investigate the affected proteins by tumor necrosis factor (TNF)-? in endothelial cells, and further explore the potential molecular mechanism of TNF-? on endothelial cells. METHODS: Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by a NO assay kit. Proteomic alterations were analyzed using two-dimensional electrophoresis, and peptide mass fingerprinting with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. RESULTS: NO production in HUVECs decreased significantly after TNF-? treatement. Proteomics analysis showed 21 protein spots were changed including 9 spots that were increased and 11 spots that were decreased after TNF-? stimulation, and 1 spot was only detected in TNF-? activated cell gels. CONCLUSIONS: The decreased expression of ecNOS by TNF-? might result in decrease in NO production. Up-regulated MAP/ERK kinase 3 expression might imply that TNF-? activates the expression of adhesion molecules. Cytoskeletal protein actin is also involved in TNF-? injuried HUVECs. Proteomic analysis can find some clues for identifying new potential target of TNF-?. [

AIM:ToinvestigatetheeffectofP21-activatedkinase1(PAK1)andlymphoidenhancer-binding factor 1(LEF1) on the proliferation of esophagus cancer cells .METHODS:Real-time PCR was applied to detect the mR-NA expression of PAK1 and LEF1 in the esophagus cancer tissues .MTT assay were used to measure the proliferation of hu-man esophagus cancer cell line KYSE transfected with PAK 1 and LEF1.RESULTS: The mRNA expression of PAK1 in the esophagus cancer tissues was lower than that in control group (P<0.05).The mRNA expression of LEF1 and tran-scription factor 4 (TCF4) in the esophagus cancer tissues was higher than that in control group (P<0.05).The prolifera-tion of KYSE cells with over-expression of PAK1 and LEF1 was higher than that in control group .No significant change of apoptosis between the KYSE cells with over-expression of PAK1 and LEF1 and control group was observed .CONCLU-SION:The expression of PAK1 decreases and the expression of LEF 1 increases in esophagus cancer tissues .LEF1 domi-nantly regulates the proliferation of esophagus cancer cells .

Objective To analyze the curative effect of minimally invasive incision combined with plate fixation for calcaneal fractures.Methods Retrospectively analyzed the clinical data of 30 cases(30 feet) of calcaneal fractures who were treated with minimal incision(sinus tarsi approach combined with posterior lateral incision) from August 2014 to August 2015.Minimally invasive incision combined with plate fixation were performed for the treatment.X-rays were taken in the regular follow-up,B(o)hler and Gissane angle as well as the complications were measured.The final curative effect was comprehensively assessed according to the ankle hind foot score of American Aryhopaedic Foot and Ankle Society(AOFAS).Results All cases were followed up for an average of 14.5 months (from 12 to 16 months), and all fractures healed with no complication such as incision infection or poor healing.The average B(o)hler angle was improved from preoperative(13.42°±4.2°) to (29.3°±3.8°) at the last follow-up with a significant difference(t=-20.424,P=0.000),and the average Gissane angle was significantly improved from preoperative(97.8°±9.7°) to (123.6°±6.5°) at the last follow-up (t=-17.5,P=0.000).The outcomes were evaluated with AOFAS,resulting in an excellent and good rate of 83.3%.Conclusion Treatment of calcaneal fractures with minimal incision has the advantages of less trauma,lower incision complication rate,stronger reduction and fixation,which can achieve satisfactory outcomes.

OBJECTIVETo evaluate the clinical effect of Dusuqing (DSQ) in treating senile bacterial pneumonia and to explore its possible mechanism.

METHODSPatients were randomly divided into two groups, the treated group treated with conventional western therapy (CWT) and the combined treated group treated with CWT plus DSQ respectively. The changes of serum cytokines, interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha), before and after treatment were observed.

RESULTSIn the 33 patients of the combined treated group, the total effective rate being 93.9%. While in the 31 patients of treated group, the total effective rate being 83.9%. The difference of the total effective rate between the two groups was significant (P < 0.05). The scores of principal symptoms, secondary symptoms and total scores were all obviously lowered in both groups, particularly in the combined treated group. The peripheral blood IL-2 level was lower and TNF-alpha higher in senile patients than those in healthy aged subjects respectively. All the two parameters approached normal level in the combined treated group but with insignificant change in the CWT treated group.

CONCLUSIONDSQ showed obvious effect in treating senile bacterial pneumonia. The mechanism might be related with the lowering of inflammatory cytokines such as oversecreted TNF-alpha, as well as the improvement of immune function (elevation of IL-2) of organism.

Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-2 ; blood ; Male ; Middle Aged ; Phytotherapy ; Pneumonia, Bacterial ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUND: The cell growth, proliferation and differentiation caused by p38 mitogen-activated protein kinase (p38MAPK) might act as the common pathway in the onset and development of diabetic vascular complication.OBJECTIVE: To investigate the effect of atorvastatin on p38MAPK signal pathway and the influence of atorvastatin on cell proliferation and expression of transforming growth factor-β1 (TGF-β1) at transcriptional level in human glomerular mesangial cells (HGMCs) cultured with oxidative modification of low-density lipoprotein (ox-LDL).DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Endocrinology Department, First Hospital Affiliated to China Medical University; Endocrinology Department,Respiratory Department, Urology Department, the General Hospital of Shenyang Military Area Command of Chinese PLA.MATERIALS: The experiment had been done in the laboratories for Pharmaceutical Department of China Medical University and Respiratory Department of Shenyang Military Area Command of Chinese PLA from May 2004 to May 2005. The sample was cut from renal cortex from the healthy segment of nephroectomy from a tumor patient (Provided by Xiang Jun, Urology Department, the General Hospital of Shenyang Military Area Command of Chinese PLA; Informed consent was obtained). OX-LDL was purchased from biochemistry institute of Peking Union Medical College (Batch No.20040711 ). ox-LDL was 5.3±1.0 nmol in 100 μg protein. Atorvastatin was purchased from Pfizer Pharmaceutical Co. Ltd (No. 45837088); p38MAPK monocloncal antibody was purchased from Santa Cruz.METHODS: ① 6.0-8.0 cm3 blocks of cortex were cut from renal cortex from the healthy segment of nephroectomy from a tumor patient, glomerular mesangial cells were isolated. When grew into 80% confluent monolayer, the cells were digested and performed passage. After the first two passages, the cells were pure based on morphology and characterized by DAB staining for Vimentin antigen and actin antigen was positive, whereas cytokeratin antigen was negative. Oil red "O" staining confirmed that ox-LDL was intaken by HGMCs. The 4-8th passages of cells were used to study. ②HGMCs were seeded into 96-well plates with 5×103 cells per well and grown in 200 μL culture medium. The study was divided into 5 groups (6 wells each group): 1.2, 6,12 mg/L atorvastatin group, ox-LDL group and blank control group. The cells were pre-incubated with atorvastatin for 30 minutes, then exposed to 80 mg/L ox-LDL. The cells in blank control group were untouched. After 24 hours, MTT was added. The absorbance of each sample at the wavelength of 492 nm was measured with immunosorbant assay system. The inhibitory rate of cell proliferation was calculated. ③1×106 to 3×106 cells were seeded into six 200 mL flasks. The trial was divided into 5 groups randomly:control group, 10, 40, 80 mg/L ox-LDL groups and atorvastatin group (12 mL/g). The cells in each group were pre-incubated for 30 minutes, then exposed to 80 mg/L ox-LDL for 24-routine culture. The expressions of TGF-β1mRNA of harvested cells were detected with semi-quantitative reverse transcription-polymerase chain reaction and p38MAPK signal pathway activation was detected by Western blot.MAIN OUTCOME MEASURES: ①Identification results of HGMCs. ② Proliferation of HGMCs. ③ TGF-β1 expression of HGMCs. ④p38MAPK signal pathway activation of HGMCs.RESULTS: ①When the cells were sub-cultured to the second generation, cell volume was big. Most of the cells were spindle-shaped, irregular stellate or branch-like, filled with microfilaments which paralleled axis. Cells overlapped in the intensive area. After DAB staining, cytoplastic actin and vimentin were positive and keratin was negative. Oil red "O"staining confirmed that ox-LDL was intaken by HGMCs with red granules in the cytoplasma, while control group did not.It was proved that the cells cultured for passage were HGMCs. ② As compared with control group, the inhibitory rate of cell proliferation in ox-LDL group was significantly decreased, but that in atorvastatin 1.2, 6 and 12 mg/L groups was significantly increased (0, -17.4%, 6.4%, 22.5%, 61.5%, respectively, P ＜ 0.05 or 0.01) on concentration-dependent manner. ③ As compared with control group, ox-LDL (10, 40, 80 mg/L) increased the expression of TGF-β1 and activation of p38MAPK in concentration-dependent manner, the effect of 80 mg/L ox-LDL group was the most significantly (P ＜ 0.01). Atorvastatin decreased the increment of TGF-β1 expression and the activation of p38MAPK pathway induced by ox-LDL significantly. There was significant difference when compared with 80 mg/L ox-LDL group (P ＜ 0.01).CONCLUSION: Atorvastatin can antagonize the activation of p38MAPK pathway, decrease the secretion of TGF-β1 and inhibit mesangial cell proliferation induced by ox-LDL, suggesting that it may exert beneficial effect in the prevention and treatment of diabetic nephropathy with dyslipidemia.