In this paper, a qualitative and quantitative analysis method was established for the scopoletin in herba Artemisiae hedinii hy HPLC. The chromatographic column was Agilent TC-C18 (4.6 mm×250 mm, 5μm);the tem-perature of column was 30℃, eluted with a mobile phase consisted of methanol and 0.3%phosphoric acid solution and gradient elution at a flow rate of 1.0 mL·min-1. The detection wavelength was 344 nm. The results showed that the mass concentration and peak area of scopoletin had a good linear relationship within the range of measurement (r=0.999 9). The reproducibility was good;and the RSD was 1.51%. The average sample recovery rate was 101.42%. It was concluded that the established method was stable, reliable and simple. It provided theoretical evidences for the quality evaluation and quality control of herba A. hedinii.

Background Heparanase degrade heparan sulfate side chains of heparan sulfate proteoglycans in the extracellular matrix.Heparanase induces angiogenesis and likely promotes the vascularization of tumor.ObjectiveThe present study is to investigate the expression of heparanase and perlecan in retinas with oxygen-induced retinopathy.Methods Sixty-five clean neonatal C57BL/6J mice were raised in a hyperbaric oxygen box with a volume percentage of 75%±2% for 5 days and then returned to the normal air room.Another 65 matched mice were raised in the normal environment as controls.Evans blue was infused by the superior vena cava in all the mice on postnatal days 12,13,17,21 and 30,afterwards fluorescein angiography was performed and then the mice were sacrificed.The retinas of mice were isolated and prepared and the retinal vessels were examined under a fluorescent microscope and optical microscope.Heranase and perlecan mRNA was detected using reverse transcription PCR (RT-PCR).Heranase and perlecan proteins were detected by Western blot.The analysis of variance was used to compare the mRNA and the protein levels of heranase and perlecan between the experimental and control groups.Results The expression of heparanase mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=16.303,P=0.000;F_(time)=18.614,P=0.000;F_(interaction)=11.299,P=0.000),and the expression of heparanase mRNA was significantly enhanced in mice from postnatal days 12,13,17 and 21 compared with normal control mice (P=0.001,0.000,0.000,0.001,respectively).The expression of heparanase protein in the retinas of different ages of mice and the different groups followed the same tendency(F_(group)=458.134,P=0.000;F_(time)=78.466,P=0.000;F_(interaction)=71.398,P=0.000).The expression of perlecan mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=7.351,P=0.013;F_(time)=9.098,P=0.000;F_(interaction)=3.349,P=0.000),and increase in differences also were clearly seen in mice from postnatal days 13,17 and 21 compared with normal control mice (P=0.048,0.000,0.003,respectively).Conclusion The expression of heparanase and perlecan is associated with the development and progression of retinal neovascularization,and perlecan and heparanase together produce a synergistic effect.Heparanase and perlecan may participate in the angiogenesis of oxygen-induced retinopathy.

OBJECTIVETo explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method.

METHODSThe retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05).

CONCLUSIONSAGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

Animals ; Cells, Cultured ; Diabetic Retinopathy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ependymoglial Cells ; Glucose ; L-Lactate Dehydrogenase ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A

OBJECTIVETo investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on activity of purified retinal ganglion cells (RGCs) cultured in high glucose medium.

METHODSPurified RGCs of SD rats were cultured in stimulative stable high glucose (50 mmol/L) condition (SHG) and fluctuated glucose condition (FGC) separately, they were intervened with S-NSAB, and the lactate dehydrogenase (LDH) leakage was detected by spectrophotometer for estimating the activity of RGCs.

RESULTSLDH leakage (U/L) in SHG culture was 1 349.17 +/- 215.50 at 24 h, 1220.24 +/- 124.53 at 48 h and 1982.14 +/- 219.03 at 72 h, all significantly lower than that in normal control at the corresponding time points (1628.10 +/-122.10, 1484.13 +/- 127.55 and 2155.75 +/- 140.44, respectively, P<0.05), whereas it was obviously higher in FGC culture at 72 h (2299.60 +/- 88.35), showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points (P<0.05). The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h (1797.62 +/- 146.40) and in FGC at 48 h (1259.92 +/- 87.74) and 72 h (1940.40 +/- 155.47), the difference between pre- and post-intervention was significant (P<0.05).

CONCLUSIONFluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells; S-NSAB could improve these abnormalities in either SHG or FGC condition, which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.

Animals ; Animals, Newborn ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; drug effects ; Serum

OBJECTIVETo investigate the clinical application of free superficial iliac circumflex artery skin flaps, as well as the management of donor site defects.

METHODS17 free superficial iliac circumflex artery skin flaps were applied for the traumatic defects or deformities on face, neck, foot, hand, ankle and lower leg, respectively. The donor site defects were closed directly or covered by paraumbilical island flaps.

RESULTSThe 17 flap size ranged from 5 cm x 3 cm to 19 cm x 14 cm. 16 flaps survived completely except 1 flap with partial necrosis, which was closed by free skin graft. The donor site defects were closed directly in 10 cases, and covered by paraumbilical island flaps in 7 flaps without no flap necrosis. The abdomen had a good appearance.

CONCLUSIONSGood appearance can be achieved with free superficial iliac circumflex artery skin flaps for the defects on face, neck, foot, hand, ankle and lower leg. Paraumbilical island flap can be used for the donor site defects.

Arteries ; Foot ; Free Tissue Flaps ; blood supply ; transplantation ; Humans ; Reconstructive Surgical Procedures ; Skin ; Skin Transplantation ; Transplant Donor Site ; surgery ; Wounds and Injuries ; surgery

OBJECTIVETo investigate the impacts of steady high-glucose or fluctuated glucose conditions on glutamate (Glu) release in purified retinal ganglion cells (RGCs) cultured in vitro, and the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on it.

METHODSRGCs of neonatal SD rats were cultured by antibody combined two-step purified method in different conditions: the simulated normal condition, the steady high-glucose condition and the fluctuated glucose condition, and they were intervened with S-NSAB. Thereby, the experiment was carried out in 6 groups, i.e. the normal control group (A), the S-NSAB intervened group (B), the steady high-glucose cultured group (C), the steady high-glucose cultured and S-NSAB intervened group (D), the fluctuated glucose cultured group (E), and the fluctuated glucose cultured and S-NSAB intervened group (F). Content of Glu in the extracellular fluid was detected at 24, 48 and 72 h after intervention with a full-automatic biochemical analyzer. And the data obtained were statistically analyzed with SPSS 13.0 soft ware.

RESULTSRelease of Glu at 24 h after intervention in Group E (256.33 +/- 25.73 mg/L) was obviously higher than that in Group A and Group C (134.22 +/- 9.14 mg/L and 141. 17 +/- 22.13 mg/L, P < 0.05); at 24 h and 72 h in Group B (124.50 +/- 10.30 mg/L and 30. 17 +/- 2.97 mg/L) was obviously lower than in Group A respectively (P < 0.05); in Group D at 24 h (127.50 +/- 16.94 mg/L), 48 h (26.17 +/- 3.99 mg/L) and 72 h (27.67 +/- 3.49 mg/L) were lower than in Group C; in Group F at 24 h (228.33 +/- 18.41 mg/L) and 72 h (28.00 +/- 2.41 mg/L) were lower than in Group E respectively at the corresponding time points.

CONCLUSIONSFluctuated glucose condition could obviously increase the Glu release of RGCs, to cause extracellular large amount Glu accumulation, which induces the exciting neurotoxicity to RGCs and finally to aggravate the injury on cells. S-NSAB could reduce the Glu release to some extent in the steady-high or fluctuated glucose conditions, diminish the injury of RGCs from exciting neurotoxicity of Glu, and it might be one of the intervening pathways of Chinese drugs for NSAB in preventing and treating DRP.

Animals ; Animals, Newborn ; Cells, Cultured ; Diabetic Retinopathy ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; pharmacology ; Glutamic Acid ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; metabolism

Objective By analyzing the urinary calculi ingredient and 24-hour urinary stone risk factors of urolithiasis patients in Hengyang area,to investigate the effects of diet intervention on recurrence rate in urolithiasis patients,and provide the measures for prevention and treatment.Methods Prospectively collected 692 patients that permanent residents in Hengyang area from September 2008 to September 2012,who had implementation of minimally invasive operation and taken stone specimens to analyze composition,and also collected 24 hours urine to analyze the urinary stone risk factors.They were divided into test group and control group by random number table method,346 cases in each,control group without diet intervention,and test group was given diet intervention according to the stone composition and urinary stonerisk factors.All patients were followed up for 1 year,the urinary stone recurrence rate in Hengyang area was observed.Results Among 692 urolithiasis patients,663 patients completed the study (test group of 341 cases and control group of 322 cases),the expulsion rate was 4.19％(29/692).The 24-hour urinary stone risk factors in control group before and after diet intervention had no significant difference(P ＞ 0.05).In test group after diet intervention,the excretion of ingredients in urine such as dietary calcium (t =3.412,P ＜ 0.05),oxalate(t =3.018,P ＜ 0.05) and uric acid(t =1.990,P ＜ 0.05) was obviously decreased,and urinary citrate (t =3.174,P ＜ 0.05) was increased,but the excretion of ingredients such as magnesium and phosphorus had no significant difference (P ＞ 0.05).After 1 year after diet intervention,the recurrence rate in test group was lower than that in control group [0.88％ (3/341) vs.7.76％ (25/322)],there was significant difference (P ＜ 0.01).Conclusion Diet intervention can effectively reduce the risk of urinary stone according to the stone composition and the 24 hours urine stone risk factors,plays an important role on reducing urinary stone recurrence,which is worth clinical promotion.

OBJECTIVE To investigate the prevalence of the antimicrobial and disinfectant-resistant genes from the clinical isolates of meticillin-resistant Staphylococcus aureus(MRSA) and meticillin-resistant Staphylococcus haemolyticus(MRSH).METHODS The identification and antimicrobial susceptibility test of MRSA and MRSH isolates were determined by MicroScan auto SCAN4.PCR was used for detecting antimicrobial and disinfectant-resistant genes.The minimal inhibitory concentration(MIC)and minimal bactericidal concentration(MBC)of benzalkonium bromide to staphylococci that carried qacA/B gene were determined by broth dilution method.RESULTS The positive rates of qacA/B,aac(6′)/aph(2″),aph(3′)-Ⅲ,ant(4′,4″),TEM,erm,and tetM from 24 MRSH strains were 37.5%,87.5%,33.3%,29.2%,95.8%,and 94.4%;and 91.7%.respectively,and 30.6%,91.7%,72.2%,8.3%,100%,94.4% and 91.7%,respectively.The MIC of benzalkonium bromide for MRSH and MRSA were both 32-128 mg/L,MBC for MRSH was 256-512 mg/L and for MRSA was 512-1024 mg/L.The MIC and MBC of benzalkonium bromide for standard strain ATCC25923 were 16 mg/L and 32mg/L,respectively.CONCLUSIONS Antimicrobial and disinfectant-resistant genes are commonly prevalent in MRSA and MRSH isolates.

Adolescent ; Adult ; Anemia, Aplastic ; etiology ; therapy ; Benzene ; poisoning ; Bone Marrow Cells ; drug effects ; pathology ; Female ; Humans ; Male ; Occupational Diseases ; etiology ; therapy ; Occupational Exposure ; adverse effects

Orjective:To obtain recombinant CHO-K1 with expressing sPDGFR? and to identify the biological activities of sPDGFR? secreted in non-serum medium.Methods:Recombinant human sPDGFR? expression vector pIRES-Neo3-sPDGFR?-Fc was constructed and then transfected into CHO-K1 cells by using LipofectamineTM 2000.After screened with G418 in 8 weeks,some monoclone cells were selected randomly to amplify in 96-well-plate to 24-well-plates,and then to identify positive cell clones by RT-PCR.Furthermore,the candidate cell clones were test by Real-Time PCR and Western blot assays.Finally,anti-proliferation activities of the expressed sPDGFR? were analyzed by MTT.Results:sPDGFR?-Fc was cloned into pIRES-Neo3 correctly.The sPDGFR?-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFR?-Fc obtained from cultured non-serum medium of positive CHO-K1 could significantly inhibit proliferation of vascular endothelial cell.Conclusion:Successed to select recombinant CHO-K1 cell lines with high expressed sPDGFR?-Fc.The sPDGFR?-Fc can inhibit the cell proliferation significantly and it means sPDGFR?-Fc might be a new anti-cancer drug in the future.