OBJECTIVE To identify the chemical constituents of Rhamni Songoricae Fructus and to establish their fingerprints and the method for simultaneous determination of four constituents to comprehensively evaluate the quality of Rhamni Songoricae Fructus. METHODS The chemical constituents in Rhamni Songoricae Fructus were qualitatively analyzed by ultra-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry. The fingerprints of 15 batches of Rhamni Songoricae Fructus were established by HPLC and chemometric analysis was performed by using SPSS 26.0 and SIMCA 14.1 software； the contents of quercetin， kaempferol， kaempferide and emodin were determined by the same method. RESULTS A total of 35 constituents were identified， including 28 kinds of flavonoids， 5 kinds of anthraquinones and 2 kinds of organic acids. A total of 19 common peaks were identified in the HPLC fingerprints， recognizing quercetin， kaempferol， kaempferide and emodin. The similarities between HPLC fingerprints of 15 batches of samples and control chromatograms were greater than 0.9. The results of cluster analysis showed that 15 batches of samples were divided into 2 classes， of which S1-S5， S7 and S9 were one class and the rest were one class， similar to the results of principal component analysis. The results of the orthogonal partial least squares- discriminant analysis showed that the variable importance projections of peaks No. 2， 6， 1， 11 （quercetin）， 3， 14， 8， 10， 19 （emodin）， 5 were all greater than 1. The contents of quercetin， kaempferol， kaempferide and emodin ranged from 0.710 to 10.478 mg/g， 0.236 to 0.660 mg/g， 0.334 to 3.039 mg/g， and 0.261 to 0.504 mg/g. CONCLUSIONS The constructed chemical constituent identification， fingerprint and content determination methods are simple， feasible and reproducible， which combined with chemometric analysis can be used for comprehensive evaluation of the quality of Rhamni Songoricae Fructus.

Objective:To investigate the effect of sorafenib on the proliferation of human oral cancer TCA8113 cells and to explore the underlying mechanisms.Methods:Mter treated with sorafenib at 2.5,5,10,20 μg/ml respectively for48 h,TCA8113 cell proliferation was examined by MTT and colony formation assay.Western blotting was employed to examine the p38MAPK expression in the cells.TCA8113 cells were pretreated with 10 μmol/L of SB203580 (a specific inhibitor of p38MAPK) for 30 min,and then by different concentrations of sorafenib for 48 h,cell proliferation was tested by MTT assay.Results:Sorafenib significantly inhibited the proliferation of TCA8113 cells in a concentration dependent fashion.Sorafenib also remarkably promoted the activation of p38MAPK of the cells.SB203580 significantly alleviated soiafenib induced TCA8113 cell viability decrease.Conclusion:Sorafenib can inhibit the proliferation of TCA8113 cells,which may be related to the activation of p38MAPK.

Objective To investigate the effect of cannabinoid receptor 1 ( CBR1 ) on spatial learning and memory function of neuropathic pain ( NP ) model rats and the expression of N-methyl-D-aspartic acid receptor 1(NR1) subunit in medial prefrontal cortex (mPFC).Methods Thirty-six healthy male Wistar rats were randomly divided into 4 groups, with 9 rats in each group: the sham operated group (SO group), the neuropathic pain model group (NP group), the NP model group with an mPFC injection of saline ( NS group ) , and the NP model group with an mPFC injection of the CBR 1 antagonist AM251 ( AM251 group).The NP model was prepared using the operation of chronic constriction injury ( CCI) of the right sciatic nerve.The mechanical withdrawal threshold ( MWT ) and the thermal withdrawal latency (TWL) of the rats in each group were detected at 3, 7, 14, 21 and 28 days after the operation.At 29 days after the operation , 18 rats of NP model were randomly selected and given an mPFC injection of saline or AM251 using a three-dimensional brain puncture.At days 30-37 after operation , the eight-arm maze test was performed to detect the spatial learning and memory function of the rats , and the rats were sacrificed immediately after this test.The expression levels of CBR1, NR1 and phosphorylated-N-methyl-D-aspartic acid receptor 1 ( p-NR1 ) ( Ser896 ) in the mPFC brain region were detected by Western blotting , RT-PCR and immunofluorescence.Results Compared with the SO group , the pain thresholds and the spatial learning and memory function of the rats in the NP group were significantly lower ( both P <0.05 ).Compared with the NS group , the rats in the AM251 group showed improvement about spatial learning and memory function ( P<0.05).Compared with the SO group ( the mRNA and protein level of CBR 1:0.23 ± 0.06,0.42 ±0.03), the mRNA(0.43 ±0.12) and protein (0.53 ±0.05) level of CBR1 in NP group increased (both P<0.05).Compared with the NS group (the mRNA and protein level of CBR1:0.42 ± 0.11,0.52 ±0.10), the mRNA (0.53 ±0.05) and protein (0.98 ±0.17) level of CBR1 in AM251 group increased (both P<0.05).Compared with the SO group (the mRNA and protein level of NR1 and the protein level of p-NR1:1.50 ±0.15,0.65 ±0.05,0.79 ±0.15), the mRNA (0.94 ±0.07) and protein (0.24 ±0.05) level of NR1 in NP group decreased (both P<0.05), the protein level of p-NR1 (0.33 ± 0.04) decreased (P<0.05).Compared with the NS group (the mRNA and protein level of NR1 and the protein level of p-NR1:1.09 ±0.14,0.26 ±0.06,0.31 ±0.08), the mRNA(1.58 ±0.10) and protein (1.42 ±0.10) level of NR1 in AM251 group increased (both P<0.05), the protein (0.95 ±0.15) level of p-NR1 increased ( P<0.05).Conclusion CBR1 can decrease the expression level of NR 1 and p-NR1 in the mPFC brain region of NP model rats and induce the spatial learning and memory impairment.

AIM:To investigate the effect of genistein on the proliferation of human oral cancer TCA 8113 cells and to explore the underlying mechanisms .METHODS:The cell proliferation was examined by MTT assay , cell counting and colony formation assay .Western blotting was employed to examine the protein levels of vascular endothelial growth fac -tor (VEGF), extracellular signal-regulated kinase (ERK) and p-ERK.RESULTS: Genistein significantly inhibited the proliferation of TCA8113 cells in a concentration-dependent fashion .Moreover , genistein dose-dependently decreased the protein levels of VEGF, ERK and p-ERK.The expression of VEGF was also blunted by U 0126, a specific inhibitor of ERK.U0126 and axitinib, a VEGF receptor antagonist , both significantly inhibited the proliferation of TCA 8113 cells. CONCLUSION:Genistein inhibits the proliferation of TCA8113 cells, which may be related to its inhibitory effect on ERK expression and activation , thus subsequently decreasing the expression of VEGF .

OBJECTIVE:To study the bioequivalence of Levodopa micro-capsule floating tablets in Beagle dogs after multi-dose administration. METHODS:6 dogs were collected and divided into Levodopa micro-capsule floating tablets group and Com-pound levodopa preparation group (Benserazide tablet,reference preparation). They were given levodopa 200 mg intragastrically, every 8 h,for consecutive 4 day. In two-period crossover test,HPLC method was established to determine the concentration of le-vodopa in dog. The pharmacokinetic parameter,bioequivalence and plasma concentration fluctuation of steady state were calculated. RESULTS:The main pharmacokinetic parameters of Levodopa micro-capsule floating tablets and reference preparation were as that cmax were(4.23±0.75)and(8.47±1.18)μg/ml;AUC0-∞ were(12.18±1.16)and(13.81±2.12)μg·h/ml;tmax were(1.83±0.26) and(0.67±0.13)h,respectively. 90% confidence intervals for the geometric mean ratio of AUC0-∞ for test and reference prepara-tion were 80.61%-97.90%,and that for cmax were 42.75%-57.63%,respectively. There was statistical significance in tmax between test and reference preparation. Degree of fluctuation of test and reference preparation at steady state were(283.914±43.217)% and (506.489±78.965)%,and fluctuation coefficient were(177.463±7.873)% and(187.405±1.650)%,respectively. The degree of fluctuation of test preparation was significantly less than that of reference preparation. CONCLUSIONS:Levodopa micro-capsule floating tablets show good sustained-release property,and are bioequivalent with reference preparation in absorption after multiple dose administration. It also has lesser fluctuation of blood concentration.

OBJECTIVE: To investigate the clinical application value of cone-beam CT (CBCT) online correcting technology in volume modulated radiation therapy (VMRT) for nasopharyngeal carcinoma (NPC). METHOD: Fifty NPC patients applying head neck and shoulder thermoplastic body membrane fixing device were eligible for treatment VMRT, these patients would accept a couple of CBCT scanning by on board imager (OBI) in a fixed time each week after initial setup and after online correcting during the first three week for radiotherapy, CBCT images and DRR images constructed by CT simulation were carried out registration,which could calculate the setup errors of initial setup and after online correcting. RESULT: Fifty patients were accepted 150 scanning after initial setup and 150 scanning after online correcting respectively, the errors after initial setup were (-1.24 ± 1.25)mm in X direction, (1.19 ± 1.85)mm in Y direction, (1.49 ± 1.70) mm in Z direction. The setup errors after online correcting were (-0.13 ± 0.29)mm in X direction, (0.10 ± 0.47)mm in Y direction, (0.17 ± 0.36)mm in Z direction. The setup errors after online correcting were significantly lower than the errors after initial setup in X direction (P < 0.05), Y direction (P < 0.05), Z direction(P < 0.05). The M(PTV) value after online correcting were 0.46 mm, 0.53 mm and 0.59 mm in X, Y and Z directions respectively. CONCLUSION The use of CBCT online correcting technology can significantly reduce the setup errors of VMRT for NPC and improve the treatment effect.
Carcinoma ; Cone-Beam Computed Tomography ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; radiotherapy ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated

Objective:To establish the quality standard for peganum harmala alkaloids cream ( CAPH) . Methods: The general quality of CAPH was inspected according to the general notices described in Chinese Pharmacopoeia volumeⅠ2010 edition. The qual-itative identification was carried out by TLC with harmine and harmaline as the index ingredients. The content determination was carried out by HPLC methods with harmine, harmaline and vasicine as the index ingredients. Results:The inspection items were all met the requirements. The experimental samples and the reference substances in TLC showed the identical spots with the same color and shape at the same position. The calibration curve of harmaline, harmine and vasicine was linear within the concentration range of 3. 440-110. 000 μg·ml-1 , 3. 340-107. 000 μg·ml-1 and 1. 380-22. 000 μg·ml-1 , respectively. The recovery was 98. 1%, 99. 8% and 99. 3% with RSD of 1. 75%, 1. 78% and 1. 95%, respectively (n=6). Conclusion: The established quality control methods meet the requirements of methodology, and the results lay foundation for the quality standard for CAPH.

AIM: To explore the effect of L-carnitine on nuclear factor of activated T-cells, cytoplasmic 3 (NFATc3) in cardiomyocytes under H2O2 stimulation.METHODS: Primary cultured neonatal rat myocardial cells were stimulated by H2 O2 at concentration of 200μmol/L for 12 h to induce oxidative stress injury.In treatment group, L-carni-tine and cyclosporin A ( CsA) , a specific inhibitor of calcineurin ( CaN) , were administered 30 min prior to H2 O2 stimula-tion.After treatment, total, cytoplasmic and nuclear NFATc3 protein levels were determined by Western blotting.The method of immunofluoresence was used to evaluate the distribution of NFATc3.RESULTS: H2 O2 treatment produced no effect on the expression of total NFATc3, but caused its translocation from the cytosolic to nuclear compartment, which was greatly blunted by L-carnitine pretreatment.CONCLUSION:L-carnitine antagonized oxidative stress injury via alleviating NFATc3 nuclear translocation.

Objective To investigate the clinical characteristics and prognosis of patients with different molecular subtypes of breast cancer.Methods A cohort of 716 breast cancer patients which had clear immunohistochemical detection were investiged.Their molecular subtypes were categorized as Luminal A,Luminal B,HER-2 over-expressing and basal-like subtypes,based on detection of ER,PR,HER-2 expression,and the clinical data including characteristics,relapse,prognosis and prognostic factors of the patients with different subtypes of breast cancer were analyzed retrospectively.Results There were no significant differences among different molecular subtypes at the age,menopausal status,production times,clinical stage,and radiation therapy(P ＞0.05).There were significant differences among different molecular subtypes at axillary lymph node starus (x2 =17.208,P =0.001),turner size (x2 =20.528,P =0.000) and operation method (x2 =24.242,P =0.000) and chemotherapy regimens (x2 =10.711,P =0.013).Univariate and multivariate analyses showed that clinical stage (x2 =17.005,P =0.002),axillary lymph node status (x2 =11.267,P =0.000) and molecular typing(x2 =125.634,P =0.000) were independent prognostic factors affecting long-term survisal rate.Conclusion Breast cancer patients in different subtypes have different long-term survival rate.The patients in basal-like subtype have the worst long-term survival rate.Molecular subtypes may provide important information to predict the prognosis of breast cancer.

ObjectiveTo evaluate the effect of electroencephalography(EEG) recognition rate resulted by stimulus of visual, auditory and combined perception. MethodsIn virtual traffic environment, the stimuli were designed in traffic information based on visual, auditory and audio-visual fusion. When one of the three stimuli appeared, the subjects completed starting and braking vehicle by imaginary using of right and left hands at the same time. The EEG signals were recorded and the imaginary motion-related features from C3, C4 were extracted and classified. ResultsThe best recognition rates in visual, auditory and fusion perception were 100％, 100％and 83％, respectively, while the averages of the rates were 68.8％ 、82.2％、76.9％, respectively. Conclusion The recognition rates are affected by the audio, visual and fusion stimulus and apparent individual differences exist.