This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVETo investigate the effect of triptolide (TPL) on the renal tissue of diabetic rats and its possible mechanisms.

METHODSSD rats were randomly divided into the normal control group (as the normal group), the diabetic model group (the model group), the low dose TPL treatment group (the low dose TPL group, TPL 0.2 mg/kg by gastrogavage), the high dose TPL treatment group (the high dose TPL group, TPL 0.4 mg/kg by gastrogavage). Equal volume of normal saline was given to rats in the normal group and the model group. Five rats were randomly selected from each group at week 4, 8, and 12 of the experiment to detect body weight, kidney weight, 24 h urinary albumin (24 h UAL), plasma glucose (FBG), total cholesterol (TC), total triglyeride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), white blood cell (WBC), and hemoglobin A1c (HbA1c). The mRNA and protein expression of regulated upon activation normal T-cell expressed and secreted (RANTES) in the renal tissue was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). The renal tissue was pathologically stained by HE, PAS, and Masson staining. The glomerular and renal tubular interstitial lesions were observed at each time point. The glomerular sclerosis index (GSI) was observed by PAS staining, and the renal interstitial filrosis index (RIFI) was calcutated.

RESULTSCompared with the same group at week 4, the expression of 24 h UAL, RANTES, GSI, and RIFI at week 12 significantly decreased in two TPL groups (P <0.01). Compared with the same group at week 8, the expression of 24 h UAL, RANTES, GSI, and RIFI at week 12 also significantly decreased in the two TPL groups (P <0. 05, P <0.01). Compared with the normal group, body weight and the kidney weight obviously decreased at week 4, 8, and 12 in the model group (P <0. 01); 24 h UAL, FBG, TG, TC, HbA1c, RANTES, GSI, and RIFI were obviously elevated (P <0.01). Compared with the model group, 24 h UAL, RANTES, GSI, and RIFI also decreased in the two TPL treatment groups (P <0.01). Compared with the low dose TPL group, they were attenuated in the high dose TPL group (P <0. 05, P <0. 01).

CONCLUSIONTPL could not only inhibit the over-expression of RANTES, but also improve the glomerular sclerosis and renal interstitial fibrosis in the renal tissue of diabetic rats.

Animals ; Chemokine CCL5 ; drug effects ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Diterpenes ; pharmacology ; Drugs, Chinese Herbal ; metabolism ; Epoxy Compounds ; pharmacology ; Glycated Hemoglobin A ; metabolism ; Immunosuppressive Agents ; pharmacology ; Kidney ; drug effects ; Kidney Diseases ; drug therapy ; Kidney Glomerulus ; metabolism ; Kidney Tubules ; metabolism ; Phenanthrenes ; pharmacology ; RNA, Messenger ; genetics ; Rats

OBJECTIVETo study the clinical effect and mechanism of auricular acupoint pressing (AAP) in treating sleep apnea syndrome (SAS).

METHODSForty-five patients with SAS were randomly divided in to the AAP group (30 patients) and the control group (15 patients) to observe the changes of clinical symptoms, apnea-hypopnea index (AHI), apnea index (AI), hypopnea index (HI) and minimum blood oxygen saturation (mSaO2) in night before and after treatment by multiple channel polysomnography (PSG).

RESULTSClinical symptoms were significantly alleviated in the AAP group after treatment, with improvement in various parameters monitored by PSG (P < 0.01), showing significantly reduced AHI, AI and HI and increased mSaO2 (P < 0.01). While in the control group, no improvement was found either in clinical symptom or in PSG parameters (P > 0.05). Comparison between the two groups showed significant difference (P < 0.01).

CONCLUSIONAAP is an effective treatment of SAS, it provides a facilitate, economic and safe therapy for early prevention and treatment to SAS.

Acupressure ; methods ; Acupuncture, Ear ; methods ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Seeds ; Sleep Apnea Syndromes ; therapy

Objective To investigate the myocardial damage in rats fed with corn from Keshan disease area added with bean puree. Methods Male Wistar rats were randomly divided into 3 groups according to their body weights, and fed with corn, corn from Keshan disease area added with bean puree, corn from non-endemic area. The GSH-Px activity of vena cardalis blood was examined in 1 and 3 months, rats were sacrificed after being fed for 6 months to examine the heart changes with HE stain. Results The three groups of GSH-Px activity were different in 1 and 3 months respectively(F=23.60,72.46, all P<0.01); GSH-Px activity was (181.58±22.15), (44.76±28.59)U/L in rats fed with corn, was (195.03±17.66), (30.38±3.35)U/L in those fed with corn added with bean puree from Keshan disease area, lower than the group fed with corn of non-endemic area[(340.90±95.42), (125.17±13.64)U/L, all P < 0.01]. But the difference of GSH-Px activity between simple corn group and corn adding bean puree groups of Keshan disease area was not obvious(P>0.05). Myocardial damage incidence of the three groups was 3/9,1/9,2/7. Difference among three groups did not have statistical significance(χ2=1.33, P> 0.05). Conclusions Only corn from Keshan disease area may induce myocardial damage pathology change. Adding bean puree into corn does not increase damage.

OBJECTIVETo summarize clinical application result of the thoracodorsal artery perforator pedicled flap for repair of soft tissue defects on the ipsilateral upper limb.

METHODSFrom September 2003 to May 2007, 8 patients (6 males and 2 females) with soft tissue defects on the ipsilateral upper limb underwent reconstruction with the thoracodorsal artery perforator pedicled flap. The age of patients was from 16- to 45-years-old with an average of 32 years. Of them, the recipient sites of 5 cases were located on the arm region, 3 cases on the forearm.

RESULTSThe minor superficial infection of 1 case occurred on the recipient site after operation and the wound gradually healed by daily change dressings. All the flaps had survived completely and the postoperative course was uneventful with satisfactory clinical results. Follow-up period ranged for 9-38 months after operation (mean, 19 months). There was no remarkable donor site morbidity. All cases had good appearance on recipient site.

CONCLUSIONThe thoracodorsal artery perforator pedicled flap is thin and suitable for repair of soft tissue defect on the ipsilateral upper limb.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Soft Tissue Injuries ; physiopathology ; surgery ; Surgical Flaps ; blood supply ; Thoracic Arteries ; surgery ; Upper Extremity ; blood supply ; surgery ; Young Adult

BACKGROUNDMetformin is the most widely used anti-diabetic drug in the world. An increasing body of evidence shows metformin also blocks cell cycle progression and selectively induces apoptosis via caspase activation in some breast tumor cells. Diffusion-weighted imaging (DWI) and bioluminescence imaging (BLI) have great potential in the evaluation of the early response to cancer therapies. We used DWI and BLI in evaluating the response of breast cancer to metformin.

METHODSThe luciferase-engineered human breast cancer cell line MDA-MB-231 was inoculated into the mammary fat pad of nude mice. Twelve female nude mice bearing tumors were divided into two groups. The mice in the treatment group received metformin (2 mg/ml in drinking water daily) after tumor inoculation, and the mice in the control group were offered drinking water without any drug added. We performed 7T magnetic resonance imaging and optical imaging every week. Imaging included T1- and T2-weighted imaging, DWI, and BLI. After imaging. The tumors were collected and subjected to histological analysis.

RESULTSThe mean photons/second of tumors in the treatment group was (3.00 ± 0.43)× 10(6) at day one, (1.01 ± 0.14)× 10(7) at 2 weeks, (5.79 ± 1.42)× 10(7) at 4 weeks, and (2.33 ± 0.70)× 10(7) at 8 weeks. The mean photons/second of tumors in the control group was (3.29 ± 0.59)× 10(6) at day one, (3.59 ± 0.63)× 10(7) at 2 weeks, (3.87 ± 0.56)× 10(8) at 4 weeks, and (4.12 ± 1.72)× 10(8) at 8 weeks. Compared to the control group, the treatment group showed an obvious decrease in the mean bioluminescence (photons/s) of the tumors and fewer metastases. Histological examination confirmed the presence of fewer metastases. DWI showed the apparent diffusion coefficient (ADC) value of the tumors; the mean ADC value was (0.9287 ± 0.04346)× 10(-3) mm(2)/s in the treated tumors and (0.7553 ± 0.01804)× 10(-3) mm(2)/s in the untreated tumors. The ADC value of tumors in the treatment group was significantly higher than the control tumors (P = 0.0013).

CONCLUSIONSThe growth and metastasis of MDA-MB-231 breast cancer may be inhibited by metformin. DWI and BLI have great potentials in the evaluation of the early response to metformin treatment. BLI has a high degree of sensitivity and is able to detect micrometastasis, thus can be used for identifying tumor metastasis in vivo.

Animals ; Breast Neoplasms ; drug therapy ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diffusion Magnetic Resonance Imaging ; Female ; Luminescent Measurements ; Metformin ; therapeutic use ; Mice ; Mice, Nude ; Multimodal Imaging ; Neoplasm Metastasis

In this study, the recombinant adenovirus (Ad) vector containing dual reporter gene [i.e. human transferrin receptor gene (TFRC) and firefly luciferase reporter gene] was constructed to provide a novel experimental tool for magnetic resonance (MR) and bioluminescence dual-modality molecular imaging. The cDNA of TFRC was amplified by polymerase chain reaction (PCR) and cloned into the multiple cloning site of pShuttle-CMV-CMV-Luciferase vector. After identification by Sfi I digestion and sequencing, pShuttle-TFRC-Luciferase vector and the adenoviral backbone vector (pAdeno) were subjected to homologous recombination. The correct recombinant plasmid was then transfected into 293 packaging cells to produce adenoviral particles and confirmed by PCR. After infection of human colorectal cancer LOVO cells with Ad-TFRC-Luciferase, the expressions of transferrin receptor (TfR) and luciferase protein were detected respectively by Western blotting and bioluminescence imaging in vitro. The results showed that TFRC gene was successfully inserted into the adenoviral shuttle vector carrying luciferase gene. DNA sequence analysis indicated that the TFRC gene sequence in the shuttle plasmid was exactly the same as that reported in GenBank. The recombinant plasmid was identified correct by restriction digestion. Ad-TFRC-Luciferase recombinant adenovirus was constructed successfully, and the virus titer was 1.6×10(10) pfu/mL. Forty-eight h after dual reporter gene transfection, the expressions of TfR and luciferase protein were increased significantly (P<0.01). It was concluded that the recombinant adenovirus vector with dual reporter gene was successfully established, which may be used for in vivo tracing target cells in multimodality imaging.
Adenoviridae ; genetics ; Genes, Reporter ; genetics ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Molecular Imaging ; methods

OBJECTIVETo investigate clinical effects of auricular acupoint (AA) in the treatment of analgesia during perioperative period in total hip arthroplasty.

METHODSFrom March 2008 to August 2010, 60 patients with late osteonecrosis of the femoral head were treated by total hip arthroplasty and randomly divided into auricular acupuncture (AA) group and control group, 30 patients in each group. There were 11 males and 19 females in the AA group,with an average age of (60.93 +/- 5.90) years; the patients were treated with auricular acupuncture on the point of Shenmen, Subcortex, Kidney and hip joint for four times a week. There were 12 males and 18 females in control group, with an average age of (59.87 +/- 6.21) years; while the patients without auricular acupuncture. VAS score was used to evaluat the degree of pain; Harris score was used to evaluat the function of hip joint. All patients received patient controlled analgesia pump (PCA) for 48 hours after surgery (400 ml liquids were in PCA pump, including 800 mg tramadol and 0.8 mg fentanyl). The dosage of liquids and adverse reaction of PCA pump in different time were recorded.

RESULTSThe VAS score on the 3rd, 4th, 5th and 7th day separately was (3.61 +/- 0.29), (3.59 +/- 0.30), (2.97 +/- 0.26), (2.29 +/- 0.45), and lower than control group, which separately was (4.19 +/- 0.28), (4.00 +/- 0.31), (3.15 +/- 0.29), (2.83 +/- 0.31). The dosage of PCA in AA group separately was (72.27 +/- 8.06), (60.40 +/- 8.16), (44.13 +/- 4.75), (40.40 +/- 3.69), and less than control group, which was (86.27 +/- 8.51), (73.87 +/- 8.32), (54.53 +/- 5.20), (44.67 +/- 6.31) on the time of 0-12, 12-24, 24-36 h and 36-48 h after surgery. During the using of PCA, nausea and vomiting occurred in 5 cases, less than control group (21 cases). Harris score in AA group (78.90 +/- 5.14) was higher than control group (73.37 +/- 5.99) 2 weeks after operation.

CONCLUSIONAuricular acupuncture can reduce postoperative pain, reduce the usage of analgesic and complications, such as nausea and vomiting, improve the function of hip joint after operation.

Acupuncture Analgesia ; Acupuncture, Ear ; Adult ; Aged ; Analgesia, Patient-Controlled ; Arthroplasty, Replacement, Hip ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; therapy ; Perioperative Period