Isolation of high-quality genomic DNA from dried and processed crude drug is the key for the DNA identification of Dendrobii Caulis. The DNA extract of Dendrobii Caulis was firstly compared using different method to isolate genomic DNA from dried and processed crude drug, including commercial DNA extracted kit and CTAB method. Using modified CTAB method (extracted from large samples), the genomic DNA was successfully isolated from Dendrobii Caulis, including Huangcao and Fengdou. The ITS regions were amplified using the purified DNA as template, and then cloned and sequenced. These ITS sequences were compared with data from Genbank database and our lab, 14 Dendrobium species and five similar species were identified from "Huangcao" and "Huangcao" slice, while six species and three similar species from "Fengdou" according to their sequence similarity. The study demonstrated that the dried Dendrobii Caulis could be identified using DNA molecular method, which could overcome deficiencies and limitations of traditional identification method and has a certain application prospects.
DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Dendrobium ; classification ; genetics ; Sequence Analysis, DNA

OBJECTIVETo evaluate the efficacy and safety of etanercept plus Tripterygium wilfordii polyglycoside (TWP) in elderly patients with active rheumatoid arthritis (RA).

METHODSTotally 46 elderly patients with active RA were randomly assigned to the treatment group (22 cases) and the control group (24 cases). All patients received subcutaneous injection of etanercept, 25 mg each time, twice per week. The dosage was reduced to once per week 3 months later. Patients in the treatment group took TWP Tablet (10 mg each time, three times per day), while those in the control group took methotrexate (MTX), 10 mg each time, once per week. The whole course lasted for 24 weeks. Patients' rest pain, tender joint number, swollen joint number, health assessment questionnaire (HAQ), patients' global assessment, physicians' global assessment, erythrocyte sediment rate (ESR), C reactive protein (CRP), rheumatic factor were assessed at week 0, 4, 8, 12, and 24. The curative effect was statistically evaluated by the United States Institute of Rheumatology ACR20, ACR50, and ACR70 improvement criteria. Meanwhile, any adverse event was recorded and evaluated.

RESULTSTotally 41 completed the trial, and 5 dropped off (3 in the treatment group and 2 in the control group). Compared with the control group, there was no statistical difference in ACR20, ACR50, or ACR70 in the treatment group (P > 0.05). Compared with before treatment in the same group, there was some improvement in tender joint number, swollen joint number, visual analogue scale (VAS) for patients' global assessment, VAS for physicians' global assessment, ESR, CRP, and HAQ between the two groups, showing statistical difference (P < 0.05). Compared with the control group in the same phase, there was no statistical difference in the treatment group (P > 0.05). There was no statistical difference in the occurrence of adverse events between the two groups.

CONCLUSIONSEtanercept plus TWP could achieve equivalent therapeutic effect to that of Etanercept plus MTX. The two regimens could improve clinical signs, symptoms, and QOL related to RA. They were well tolerated in the treatment of elderly patients with active RA.

Aged ; Antirheumatic Agents ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Drug Therapy, Combination ; Etanercept ; Female ; Glycosides ; therapeutic use ; Humans ; Immunoglobulin G ; therapeutic use ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Treatment Outcome ; Tripterygium ; chemistry

OBJECTIVETo compare the effect of reinforcing Shen method (RSM) and activating blood method (ABM) in treating osteoarthritis (OA) at the molecular level.

METHODSThe physical and chemical characteristics of components from respective recipes of RSM and ABM, and network features of component-target interaction network were analyzed by computer simulation methods including chemical space, molecular docking, and biological network, etc.

RESULTSThe chemical components of RSM and ABM were scarcely scattered with larger overlapping. Among established networks, the distribution of network features was partially similar in RSM and ABM. The average target number correlated with each component was 1.86 in RSM and 2.11 in ABM respectively. Each average target number was respectively correlated with 4.46 compounds and 3.93 compounds, reflecting multi-component and multi-target actions.

CONCLUSIONComputer simulation could intuitively trace out similarities and differences of two different methods and their interaction with targets, which revealed that the compatibility of RSM and ABM could have broader protein targets and potential synergism at the molecular level.

Computer Simulation ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Osteoarthritis ; drug therapy ; Phytotherapy ; methods

Objective Toanalyzetheclinicalcharacteristicsandtreatmentoutcomesofspinalfilum terminalevascularmalformation.Methods Theclinicaldataof6patientswithfilumterminalevascular malformation diagnosed and treated from January 2008 to December. 2013 were analyzed retrospectively. The definition of filum terminale vascular malformation is anterior/posterior spinal artery feeding arteriovenous fistula or arteriovenous malformation and located below conus medullaris,and does not complicate with spinal vascular lesions in the other part. The Aminoff & Logue score and MRI of spinal cord function were performedatoneyearaftermicroneurosurgeryand/orendovascularembolization.Results Allpatients were males. Their clinical presentations were the weakness of both lower extremities and sphincter disturbance. The mean course of disease was 17. 1 ± 5. 2 months. The pathological type of the 6 patients were all arteriovenous fistulas. The feeding arteries included lumbar artery,internal iliac artery,and median sacral artery. Two of the 6 patients underwent Onyx glue embolization,3 were treated with microneurosurgery,and 1 was treated with embolization in combination with microneurosurgery. They were all achieved anatomic cure. The Aminoff & Logue scores were improved after 1 year (3. 8 ± 1. 9 scores before procedure,2. 8 ± 2. 0 scores after procedure),there was no significant difference (P >0. 05). The myodynamia scores were improved in 3 patients,2 did not change,1 got worse. The urinary and bowel functions were improved in 2 patients,and4didnotchange.Conclusion Filumterminalevascularmalformationisararevascular malformation of spinal cord. Both embolization and surgical treatment can achieve anatomic cure. Although the spinal cord function can be only partially restored,but continuous deterioration can be prevented.

OBJECTIVE: To investigate a nwe, simple technique for preparation of interferon-alpha-liposomes, which may be suitable for industrial use. METHODS The uniform design coupled with computerized optimization was utilized to screen the formulation and preparation procedure of interferon-alpha-liposomes. Pro-liposomes were prepared by the powder bed grinding method and combined with interferon-alpha-solution to form interferon-alpha-liposomes. Liposome size was determined by the particle size analyzer. Free interferon-alpha and interferon-alpha-liposome were separated by gel filtration. Then the recovered activity of interferon-alpha was analyzed by enzyme-linked immunosorbent assay. RESULTS The result demonstrated that the best interferon-alpha-liposome formulation was as follows: the protectant was sorbitol; weight ratio of protectant to lipid was 5:1; weight ratio of octadecytamin to lipid was 1:9; weight ratio of sobey phosphatidylcholine to cholesterol was 9:1 respectively. Interferon-alpha-liposome size determined by the particle size analyzer was 80.8+/-36 nm and the encapsulation efficiency was 59.0+/-3.3%. CONCLUSION The powder bed grinding method can be used to prepare pro-liposomes which can be easily combined with interferon-alpha-solution to form interferon-alpha-liposomes.

Objective To investigate the clinical value of ~(99)Tc~m-ethylenedicysteine (EC) diuretic renography (DR) in pre-and post-operative pediatric congenital hydronephrosis.Methods The DR with injection of Furosemide at 15 min of forty children with hydronephrosis was retrospectively studied.The preoperative renal blood perfusion rate (BPR),effective renal plasma flow (ERPF),grade of hydronephrosis,renogram and renal dynamic imaging of pre- and post-operative kidneys were compared.The t-test and Mann-Whitney test were used for data analysis.Results (1) Of 40 pathological kidneys,the BPR increased 5.99% (t=-5.13,P＜0.01)from pre-operative to post-operative:(34.05±11.07)% to (40.04±8.56)%.The ERPF increased 12.48 ml/min(t=-4.35,P＜0.01) from pre-operative to post-operative:(57.81±34.32)ml/min to(70.29±5.37)ml/min.(2)The grade of hydronephrosis of 40 pathological kidneys improved significantly(Z=-2.64,P＜0.01) with the mean sum of ranks of 47.21 pre-operatively to 33.79 post-operatively.(3) As the hydronephrosis worsened,the collecting system became bigger,the renal parenchyma became thinner,the extent of intrarenal parenchymal photopenia became larger and the response to diuretic challenge in pathological kidneys decreased or became totally irresponsive.(4)Thirty-seven cases of obstruction at ureteropelvic junction (UPJO) and 3 cases at ureterovesical junction (UVJO) were diagnosed by DR,which were all confirmed by surgery.Conclusions DR is a reliable method to evaluate pediatric congenital hydronephrosis.It can accurately reflect the grade and (or) severity of the disease,guide therapy and assess the therapeutic success of operation.

Objective To explore genes expression of adiponectin receptors during differentiation of SW872 preadipocytes. Met-(hods) SW872 preadipocytes were cultured in vitro and induced to differentiate by 0.6 mmol/L oleic acid. During the progress of diffe-(rentiation), the morphological changes of SW872 cells were observed and the differentiation rate was assayed by oil-red O staining. Adiponectin receptors mRNA was measured by reverse transcription-polymerase chain reaction during differentiation of SW872 preadipocytes. Results 1.After stimulated by 0.6 mmol/L oleic acid for 72 hours, almost all SW872 cells were differentiated,and there were lots of fat droplets in the cells.2.There were adiponectin receptors genes expressions in SW872 preadipocytes.After 72 hours,and the levels of adiponectin receptor(AdipoR) 1 mRNA and AdipoR 2 mRNA were markedly increased up to 2.54 and 4.09 times,respectively. Conclusion There are AdipoR1 and AdipoR2 genes expressions in fat cells and the expressions are differentiation-dependent.

Objective To study the effect of oleic acid on the differentiation of SW872 preadipocytes.Methods SW872 prea-(dipocytes) were cultured and induced to differentiate by 0.6 mmol/L oleic acid in vitro.After 24 h,48 h and 72 h of differentiation,the morphological changes of SW872 preadipocytes were observed and the differentiation rate was assayed by oil-red O staining.In addition,triglyceride(TG) mass was detected by chemical colorimetry methods.During the differentiation of SW872 preadipocytes,transcription factors including peroxisome proliferator activated receptor-?_2(PPAR-?_2) and CAAT/enhancer binding protein-?(C/EBP-?)(mRNA) were also measured by reverse transcription-polymerase chain reaction(RT-PCR).Results 1.SW872 preadipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,when stimulated for 72 hby 0.6 mmol/L oleic acid,SW872 preadipocytes became more bigger and rounder and differentiated into mature adipocytes with lots of fat droplets in the cells.2.Compared with that of predifferentiation,the concentration of TG mass increased by 14 folds after 72-hour differentiation(P

Objective To explore the changes of soluble tumor necrosis factor receptor type 1(sTNFR1)in children with acute virus encephalitis(VE)and its clinical significance.Methods The levels of tumor necrosis factor-?(TNF-?),sTNFR1 and neuron specific enolase(NSE)in cerebrospinal fluid were determined by enzyme-linked immunosorbent assay(ELISA)in 55 children who were admitted with VE,including 25 cases with severe VE(SVE)and 30 cases with mild VE(MVE).Fifteen cases without VE were as control group.Results The levels of TNF-?,sTNFR1 in cerebrospinal fluid in encephalitis children were significantly higher than that in control group(Pa

Objective To establish a novel direct radio-labeling method for 99Tcm-IgG and evaluate the biologic distribution of 99Tcm-IgG.Methods IgG protein was modified with 2-mercaptoethanol.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),UV-vis spectrophotometer,matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) were used to identify the concentration,integrity of the modified protein.Then radiolabeled IgG-SH with 99TcmO4-was analyzed with radio-HPLC.The product was prepared as frozen kits.The distribution and metabolic process of 99Tcm-IgG were observed in New Zealand white rabbits.Results The relative molecular mass of IgG-SH and IgG measured by SDA-PAGE were similar.The relative molecular mass measured by MALDI-TOF was 1.47× 105.The radiolabeling yield was over 95％,the specific activity was 1.7× 105 GBq/mmol.All the radioactive conjugates of 99Tcm-IgG showed excellent stability in vitro.And more than 95％ conjugates retained their original structures for 6 h in 5％ BAS.Gamma imaging in New Zealand white rabbits showed blood retention in first 4 h after injection,and prominent uptake of radiotracers in the liver,kidneys,and urinary bladder at 24 h after injection,which indicated that 99Tcm-IgG was excreted mainly through the renal route.99Tcm-IgG kept its original biological activity after the modification.Conclusions Direct radio-labeling of 99Tcm-IgG was successfully established.The methods may be useful for radio-modification of monoclonal antibody.