Five flavone C-glycosides were isolated from the methanol extract of the degrease seeds of Ziziphus jujuba var. spinosa though various column chromatography methods including silica gel, MPLC, and HPLC. The structures were elucidated as 6"-feruloyl- 6'''-vanillylspinosin(1), 6",6'"-diferuloylspinosin(2), spinosin(3), swertisin(4) and isoswertisin(5) based on the NMR and MS spectral data. 1 is a new compound.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; Seeds ; chemistry ; Ziziphus ; chemistry

Objective To evaluate the effects of TENS on metatarsus plantar flexion and inversion in stroke patients,and to explore its mechanism.Methods Thirty-two stroke patients with gastrocnemius spasticity were randomly divided into a control group (n=16) and a TENS group (n=16).All patients were treated with foot sup- ports,neurodevelopmental and manipulation therapies.In addition,the TENS group received TENS on the anterior tibialis,peroneus longus and brevis muscles.All patients were assessed in terms of their Chinese stroke scale(CSS) and H reflex scores before and after therapy.Results Compared with those in the control group,the H reflex scores in the TENS group were obviously decreased,while H reflex latency was prolonged and H/M was reduced. Gait in the TENS group was evidently improved.Conclusion TENS is an effective therapy to decrease gastrocnemi- us spasticity and to improve the gait of stroke patients.

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Glutamine is an important conditionally necessary amino acid in human body. The effort is to establish a new and high efficient L-glutamine production system instead of traditional fermentaion. In this paper, high efficiency of L-glutamine production is obtained by coupling genetic engineered bacterial glutamine synthetase (GS) with yeast alcoholic fermentation system. Glutamine Synthetase gene (glnA) was amplified from Bacillus subtilis genomic DNA with primers designed according to sequences reported in EMBL data bank, then it was inserted into expression vector PET28b, the sequence of glnA was proved to be the same as that reported in the data bank by DNA sequencing. After transformation of this recombinant plasmid PET28b-glnA into BL-21 (DE3) strain, Lactose and IPTG were used to induce GS expression at 37 degrees C separately. Both of them can induce GS expression efficiently. The induced protein is proved to be soluble and occupies about 80% of the total proteins by SDS-PAGE analysis. The soluble GS was purified by Ni2+ chelating sepharose colum. After purification, the purified enzyme was proved active. Results reveal that the optmum temperature of this enzyme is 60 degrees C and optmum pH is 6.5 in biosynthetic reaction by using glutamate, ammonium choloride and ATP as substrates. After induction, the enzyme activity in crude extract of BL-21/PET28b-glnA is 83 times higher than that of original BL-21 extract. Mn2+ can obviously increase the activity and stability of this enzyme. Experiments show that the transformation efficiency of glutamate to glutamine is more than 95%. Because of the high cost from ATP, a system coupling GS with yeast for ATP regenaration was established. In this system, GS utilizes ATP released by yeast fermentation to synthesize L-glutamine. Yeast was treated by 2% toluence to increase its permeability and a yeast named YC001 with high yield of glutamine by coupling with recombinant GS was obtained. The good efficiency was achieved with the presence of 250 mmol/L glucose and 200 mmol/L phosphate, the transformation efficiency of glutamate to glutamine in this system is more than 80%, the average yield of glutamine is about 22g/L. This provides the basis for future large scale production of L-glutamine.
Bacillus subtilis ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; methods ; Glutamate-Ammonia Ligase ; biosynthesis ; genetics ; Glutamic Acid ; metabolism ; Glutamine ; biosynthesis ; genetics ; Yeasts ; genetics ; metabolism

OBJECTIVETo investigate the feasibility and reliability on the quantitative evaluation of Colles' fracture by multislice CT (MSCT) multiplanner reconstruction (MPR).

METHODSA total of 36 patients with Colles' fracture from July 2011 to July 2014 were investigated in this study. There were 11 males and 25 females with a mean age of (42.5 ± 5.4) years old (ranged 35 to 72 years). All the patients underwent anteroposterior and lateral X-ray films and MSCT scans on wrist joints within 2 days after trauma. Images were sent to the workstation through picture archiving and conserving system (PACS). One associate chief physician independently and respectively measured the dorsal intercalation depth of distal fracture block, palmar angle and dislocation degree of wrist articular surface collapse on anteroposterior and lateral X-ray film and MSCT-MPR. The time interval between the two measurements was 2 weeks. All the data between the first and second measurement on X-ray and MPR and the mean value between the X-ray and MPR was examined with paired t-test. The pearson analyzed their correlation.

RESULTSAmong the 35 cases, 35 cases of palmar angle, 21 cases of intercalation depth and 16 cases of dislocation of wrist articular surface collapse could be measured on both X-ray and MPR. For the above parameters, the first measurement results were (12.5 ± 3.6)°, (4.5 ± 2.1) mm, (3.7 ± 1.6) mm and the second measurement results were (4.8 ± 2.2)°, (6.4 ± 3.6) mm, (2.5 ± 1.2) mm on X-ray films respectively. The first measurement results on MPR were (14.5 ± 5.3)°, (4.2 ± 1.2) mm, (5.7 ± 2.3) mm, and the results were (13.2 ± 2.6)°, (4.7 ± 2.2) mm, (4.6 ± 2.1) mm for the second measurement respectively. The three parameters between the first and second measurement on plain film had statistical difference and low correlation (r = 0.681, 0.640, 0.345, P < 0.05). The data between the first and second measurement on MPR showed that the dislocation degree of wrist articular surface collapse had statistical difference (P < 0.05) and no statistical significance was found for the other two parameters (P > 0.05), with the moderate correlation (r = 0.954, 0.854, 0.642). The three parameters had low or moderate correlation with each other on X-ray (r = 0.454, 0.532, 0.378, P < 0.05), compared with the mean value on MPR.

CONCLUSIONUsing MSCT MPR images may carry on the multiple parameter measurement of Colles fracture, to make quantitative evaluation, and repeated measurement is better reliability.

Adult ; Aged ; Colles' Fracture ; diagnostic imaging ; Feasibility Studies ; Female ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Multidetector Computed Tomography ; methods

OBJECTIVETo study the effects of two specific cyclooxygenase inhibitors (SCI), rofecoxib and celecoxib, combined with chemotherapeutic drugs 5-Fu, DDP and VP-16 on gastric cancer cell line BGC-823, and to evaluate whether specific cyclooxygenase inhibitors can be used as a synergetic agent in chemotherapy.

METHODSThe gastric cancer cell line BGC-823 cells were incubated for 48 hours with rofecoxib and celecoxib, 5-Fu, DDP and VP-16 (concentration gradient of 5-Fu, DDP and VP-16:1 microg/ml, 10 microg/ml and 100 microg/ml), or in combination, respectively. MTT working solution was added to each culture and calculated the survival rates of gastric cancer cells. Median-effect principle and Professor Jin's evaluation methods were applied to detect the interaction between the specific cyclooxygenase inhibitors and chemotherapeutic agents.

RESULTSThe inhibition rates of gastric cancer cells were 42.63% +/- 1.26% and 50.67% +/- 2.35% by treatment with 0.1 micromol/L rofecoxib and 50 micromol/L celecoxib, respectively. The inhibition rates of gastric cancer cells by treatment with 5-Fu, DDP and VP-16 at different concentrations (1 microg/ml, 10 microg/ml and 100 microg/ml) were 39.75% +/- 3.14%, 49.96% +/- 2.08%, 87.93% +/- 3.66%; 48.28% +/- 2.08%, 59.46% +/- 1.69%, 88.23% +/- 4.81%; and 29.23% +/- 3.27%, 49.34% +/- 3.75%, 79.24% +/- 2.44%, respectively. However, the inhibition rates showed a synergetic role while combined the two SCI (0.1 micromol/L rofecoxib and 50 micromol/L celecoxib) with chemotherapeutic agent at different concentrations (P <0.05).

CONCLUSIONBoth rofecoxib and celecoxib have an ability to suppress gastric cancer cells in vitro, and the synergetic role becomes evident when rofecoxib and celecoxib are combined with chemotherapeutic agents at different concentrations, which indicate that the two specific cyclooxygenase inhibitors may be used as a chemotherapeutic sensitizer.

Adenocarcinoma ; pathology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Celecoxib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Cyclooxygenase Inhibitors ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Etoposide ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Lactones ; pharmacology ; Pyrazoles ; pharmacology ; Stomach Neoplasms ; pathology ; Sulfonamides ; pharmacology ; Sulfones ; pharmacology

OBJECTIVETo investigate the cellular effects of Pien Tze Huang (PZH) in the HT-29 human colon carcinoma cell line.

METHODSThe viability of HT-29 cells was determined by MTT assay. A fluorescence-activated cell sorting (FACS) analysis with annexin-V/propidium iodide (PI) and JC-1 staining were performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase 3 was evaluated by a colorimetric assay. The mRNA expression levels of Bcl-2 and Bax were measured by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSPZH, in a dose- and time-dependent manner, reduced viability and induced apoptosis of HT-29 cells. Moreover, PZH treatment resulted in the collapse of the mitochondrial membrane potential, activation of caspase 3, and an increase in the Bax/Bcl-2 ratio.

CONCLUSIONPZH inhibits the growth of HT-29 cells by inducing cancer cell apoptosis via regulation of the Bcl-2 family and activation of caspase 3, which may, in part, explain its anticancer activity.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Colonic Neoplasms ; enzymology ; pathology ; Drug Screening Assays, Antitumor ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Activation ; drug effects ; HT29 Cells ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC).

METHODSA immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis.

RESULTSIn 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05).

CONCLUSIONThe abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

Carcinoma, Squamous Cell ; Cyclin D1 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms ; Prognosis

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

OBJECTIVE: To reconstruct the course of vehicle collision, so that to provide the reference for forensic identification and disposal of traffic accidents. METHODS: Through analyzing evidences left both on passengers and vehicles, technique of momentum impulse combined with multi-dynamics was applied to simulate the motion and injury of passengers as well as the track of vehicles. RESULTS: Model of computer stimulation perfectly reconstructed phases of the traffic collision, which coincide with details found by forensic investigation. CONCLUSION Computer stimulation is helpful and feasible for forensic identification in traffic accidents.
Accidents, Traffic ; Automobiles ; Computer Simulation ; Forensic Medicine/methods* ; Humans ; Models, Theoretical ; Software ; Wounds and Injuries/etiology*