Objective To establish a HPLC method for determination of Chlorogenic acid in Liyan Granule. Method The determination was performed on Diamondsil Cl8 (4.6 mm?150 mm, 5 ?m) with mobile phase consisted of acetonitrile-0.4% phosphonic acid solution (13∶87) at a flow rate of 1.0 mL/min. The detective wavelength was set at 327 nm. Result The linear range of Chlorogenic acid was 0.026～0.26 ?g (r =0.999 9) and the average recovery rate of Chlorogenic acid was 101.84% (RSD=0.48%, n =6). Conclusion This method is simple, accurate and with good stabilization, and suitable for the quality control of Liyan gramule.

Whole-body PET-CT scanning has been used in the diagnosis and treatment of tumor to providing functional and morphological imaging. As the most widely used semi-quantitative analysis index, the cutoff value of SUV would affect the final diagnosis, therapeutic regimen and prognosis estimation. Therefore, the article reviews the influential factors of SUV, its correction methods, its cutoff values of differential various tumors and the progress of SUV. Although there is no acknowledged cutoff value of SUV in tumor diagnosis, SUV is still an important semi-quantitative index, and its rational and prudent use would increase both of the sensitivity and specificity in tumor diagnosis.

PET-CT, SPECT, and SPECT-CT examinations could lead to inevitable medical exposure for patients, and the radiation dose of this examinations and whether it was harmful to patients have attracted attention from the public and professionals. At the same time, there was still a controversy about whether the biological effect of low dose radiation was useful to human body. The research investigates and reviews the radiation dose of PET-CT, SPECT, and SPECT-CT scans and their biological effects. Under achieved to the demands of clinical conditions, operators should try to reduce the radiation dose of PET-CT, SPECT, SPECT-CT scans for patients. Although PET-CT, SPECT, SPECT-CT scans deliver inevitable radiation, the dose of single examination will not lead to deterministic effect, and even the view of the random effect of linear no-threshold from these scans also have been questioned.

Animals ; Bone Neoplasms ; Bone and Bones ; Humans ; Models, Animal ; Neoplasms ; Pain

Objective To observe the functional changes of T lymphocytes in the spleen of rats with bone cancer pain.Methods forty-one healthy female Sprague-Dawley rats were used in this study, and were divided into blank control, PBS and Walker-256 tumor groups.Bone cancer pain model was established by inoculation of Walker 256 cancer cells into the tibial cavity.The paw withdrawal threshold (PWT), paw withdrawal thermal latency (PWL), and spontaneous pain (SP) were all measured before modelling (as base) and at 4, 6, 8, 10, 12, 14, 16, 18, and 20 days after modelling. The function of T lymphocyte proliferation, and the content of T lymphocytes and their subgroups in the spleen were detected by cell counting kit-8 method and flow cytometry, respectively, on day 20 after modelling.Results Before modellng, there were no differences of PWT, PWL, and SP between the PBS and model groups.After modelling, the PWT and SP of model group were significantly decreased on day 4, and were always lower than that of PBS group during the experiment.Statistical analysis revealed that Walker-256 cancer cell inoculation in the tibia induced a significant decrease in PWL on day 8, 10 and 12 after modellng.Compared with the control group, T lymphocyte proliferation, content of T lymphocyte (CD3) and subgroups ( CD4 and CD8) in the PBS group were not significantly decreased.However, T lymphocyte proliferation and the content of CD3 lymphocytes in the model group were significantly lower than those in the blank control group and/or PBS group.Conclusions The bone cancer pain rat model may appear obvious mechanical allodynia and spontaneous pain.Its thermal pain hyperalgesiaonly occurred in the intermediate stage of bone cancer pain. The content of T lymphocytes and its subgroups, and the function of T lymphocyte proliferation are weakened to some extent in the bone cancer pain rat model.

OBJECTIVETo observe the intervention of electroacupuncture (EA) with different current frequencies and treatment frequencies on pain thresholt in rats with bone-cancer pain, so as to optimize treatment parameters of EA against bone cancer pain; and by measuring gene expression of opioid receptor and precursor in different tissues to preliminarily explore the possible mechanism of EA against bone cancer pain.

METHODSNinety healthy female SD rats were randomly divided into a control group, a model group, EA groups (6 subgroups according to different frequencies) and a sham EA group, ten rats in each one. Rats in the control group were injected with 10 µL of amicrobic phosphate buffer solution (PBS) into tibial cavity; rats in the remaining groups were injected with Walker 256 cancer cells to establish model of bone-cancer pain. No treatment was given to rats in the control group and model group; rats in the EA groups were treated with EA at bilateral "Housanli" (ST 36) and "Genduan" with 3 different current frequencies (2 Hz, 100 Hz and 2 Hz/100 Hz), once a day and once every other day, 30 min per treatment (1mA for 15 min, 2 mA for 15 min); rats in the sham EA group were treated with identical acupoints as the EA group, but the acupoints were needled subcutaneously and EA was connected with power off. All the treatment was given for 14 days. Dynamic plantar aesthesiometer was applied to measure the paw withdrawal thresholds (PWTs) of the affected side before the model establishment, 6d, 8d, 10d, 12d, 14d, 16d, 18d, and 20d after model establishment. The mRNA expressions of µ-opioid receptor (MOR), κ-opioid receptor (KOR), δ-opioid receptor (DOR), proopiomelanocortin (POMC) and prodynorphin (PDYN) in dorsal root ganglion (DRG) and lumbar spinal cord dorsal horn (SCDH) of L4-L6 of the affected side were detected by PCR method.

RESULTSThere were no differences in PWTs among all groups before model establishment (P>0. 05). Each time point after model establishment, PWTs in model group were obviously lower than those in the control group (all P<0. 01). Compared with the model group, PWTs in each EA subgroup were all increased (all P<0.05), but the differences at different time points were not significant among EA subgroups (P>0.05). The mRNA expressions of MOR, KOR, POMC, and PDYN in L4-L6 DRG in the 2 Hz/100 Hz II group were significantly higher than those in model group (P<0. 05, P<0. 01), while the mRNA expressions of MOR, KOR, DOR, POMC and PDYN in SCDH were not different compared with the model group (P>0. 05).

CONCLUSIONEA treatment has obvious analgesic effect on bone-cancer pain, however, its effect is not related with current frequency and treating frequency. EA against bone-cancer pain may be related with increasing the mRNA expression of some peripheral opioid receptors and precursor.

Acupuncture Analgesia ; instrumentation ; methods ; Acupuncture Points ; Animals ; Bone Neoplasms ; complications ; Electroacupuncture ; instrumentation ; methods ; Enkephalins ; metabolism ; Female ; Ganglia, Spinal ; metabolism ; Humans ; Pain ; etiology ; genetics ; metabolism ; Pain Management ; instrumentation ; methods ; Protein Precursors ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid ; genetics ; metabolism

[ ABSTRACT] AIM:To investigate the effects of tanshinone IIA ( Tan IIA) on proliferation, apoptosis and its mo-lecular mechanism in human hepatoma HepG2 cells under hypoxic condition.METHODS:Hypoxia model was established by treatment with cobalt chloride ( CoCl2 ) .The cells were divided into normoxia control group, hypoxia control group and hypoxia combined at different concentrations of Tan IIA groups.After HepG2 cells were incubated with different concentra-tions of Tan IIA (0.5, 1.0, 2.0, 5.0 and 10.0 mg/L) for 24 h, 48 h and 72 h under hypoxic condition, the cell prolifer-ation was determined by MTT assay.After Tan IIA was added to the media at different concentrations for 24 h and 48 h, the apoptotic cells were observed by Hoechst 33258 staining.The protein levels of hypoxia-inducible factor 1 alpha (HIF-1α) , vascular endothelial growth factor ( VEGF) and wild-type P53 were detected by Western blotting after cultured with different concentrations of Tan IIA for 48 h.RESULTS:Tan IIA inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner.Tan IIA induced the typical morphology of apoptotic cells and increased the apoptotic rate in a dose-and time-dependent manner after treatment with 1.0 mg/L~5.0 mg/L for 24 h and 48 h under hypoxic condition. The protein levels of HIF-1αand VEGF were weakly expressed in HepG2 cells under normoxia but up-regulated after incu-bated under hypoxia for 48 h.The protein expression of HIF-1αand VEGF were decreased with the increase in the concen-tration of Tan IIA under hypoxia.The protein expression of wild-type P53 was increased with the increase in the concentra-tions of Tan IIA under hypoxia.CONCLUSION:Tan IIA significantly inhibits the proliferation and induces the apoptosis of human hepatoma HepG2 cells under hypoxia, which may be related to the down-regulation of HIF-1αand VEGF and up-regulation of wild-type P53.

One hundred and eleven patients with acute myocardial infarction and without known diabetes mellitus who underwent continuous glucose monitoring were divided into normoglycemia(n = 30),transient hyperglycemia(n = 36),and persistent hyperglycemia(n = 45)groups.Compared with other two groups,higher mean blood glucose,standard deviation of blood glucose,largest amplitude of glycemic excursions,mean amplitude of glycemic excursions,and absolute mean of daily differences were observed in the patients with persistent hyperglycemia group(all P<0.01),who were more likely to be female with the history of hypertension and old myocardial infarction(all P<0.05).It was shown that the levels of aspartate aminotransferase,creatine phosphokinase(CK),CK-MB,total cholesterol,triglyceride,low-density lipoprotein cholesterol,HbA1C,and C reactive protein levels were higher in these patients(P<0.01).

Aim To investigate the mechanism of demethylation on adenosine (ADO )and homocysteine (HCY)-induced apoptosis in human hepatoma HepG2 cells .Methods HepG 2 cells were treated with differ-ent concentrations of ADO (1.0、2.0、4.0 mol · L-1 ) alone or in combination with HCY for 6h,12h and 24h,5-aza-2-deoxycytidine (5-Aza-CdR)as a positive control.Cell viabilities were assessed by CCK8 assay. Cell apoptosis was observed by AnnexinV-FITC/PI staining.The mitochondrial membrane potentials(ΔΨ) were measured by flow cytometry.The mRNA and pro-tein expressions of caspase-3,caspase-8,caspase-9, MDM-2,p53,Cytochrome C,DNMT1,DNMT3a,DN-MT3 b were detected by RT-qPCR and Western blot re-spectively.Results ADO alone or in combination with HCY significantly reduced the viability of HepG2 cells in a dose and time-dependent manner.The apoptotic rates of HepG2 cells after combination treatment with ADO and HCY at 1 .0,2.0,4.0 mol · L-1 for 24 h were (1 8.63 ± 1.25 )%,(29.42 ±2.37 )% and (42.47 ±3.09 )%,compared with the control group (1.30 ±0.82 )%,P <0.01;and the mitochondrial membrane potentials were decreased from 674.15 ± 82.8%(black control group)to (428.38 ±54.5)%, (297.78 ±30.5)%,(74.45 ±5.73)%,P<0.01, respectively.The expressions of caspase-3,caspase-8, caspase-9,MDM-2,p53,Cytochrome C were up-regula-ted and MDM-2 were down-regulated after combination treatment of ADO and HCY.The mRNA expressions of DNMT1 ,DNMT3 a and DNMT3 b were down-regulated after combination treatment with ADO and HCY or 5-Aza-CdR alone.Conclusion Combination treatment of ADO and HCY can cause cellular methylation chan-ges.The effects of demethylation of ADO and HCY may activate p53 gene and mitochondrial pathway, which at last leads to HepG2 cell apoptosis.

Objective To investigate the effect ofolipoprotein E (ApoE) mimetic peptide on neural apoptosis and autophagy and their mechanisms in mice after traumatic brain injury (TBI).Methods A total of 40 health adult male C57BL/6J mice were randomly divided into sham-operated group,TBI+normal saline (NS) group,TBI+COG1410 (1 mg) group and TBI+COG1410 (2 mg) group (n=10).The TBI models of moderate mice were constructed by controlled cortex impact (CCI) devices in the later three groups and mice in the sham-operated group were performed bone window operning only.Thirty min after model making,COG1410 treatment was given by intravenous injection of COGl410 via the tail vein at a dose of 1 mg/kg.d or 2 mg/kg.d.Mice in sham-operated group and TBI+NS group were injected with equal sterile NS instead.Neurological functions were tested 3 d after TBI by rolling-bar test and modified neurological severity scale (mNSS).Neural apoptosis was analyzed by TUNEL and autophagy protein LC3 expression in the neurons of cortex around the lesion focus was detected by immunofluorescence.Western blotting was employed to detect the expressions of apoptosis-related proteins (Bax,Bcl-2 and Caspase-3) and autophagy proteins (Beclin-1,LC3-Ⅰ and LC3-Ⅱ),and changes of Akt,mTOR,phosphorylated-(p-) Akt,p-mTOR levels.Results As compared with those in the sham-operated group,significantly shortened rotarod latency,significantly increased mNSS scores,significantly increased Bax and Caspase-3 protein expressions,significantly decreased Bcl-2,significantly increased Beclin-1 and LC3-Ⅱ protein expressions and number of TUNEL postive neurons,and statistically increased p-Akt and p-mTOR levels in the TBI+NS group were noted (P＜0.05).As compared with those in the TBI+NS group,significantly increased rotarod latency,significantly decreased mNSS scores,significantly decreased Bax and Caspase-3 protein expressions,significantly increased Bcl-2,significantly decreased Beclin-1 and LC3-Ⅱ protein expressions and number of TUNEL postive neurons,and statistically increased p-Akt and p-mTOR levels in the TBI+COG1410 (1 mg) group and TBI+COG 1410 (2 mg) group were noted (P＜0.05).As compared with those in the TBI+COG 1410 (1 mg) group,significantly increased rotarod latency,significantly decreased mNSS scores,significantly decreased Bax and Caspase-3 protein expressions,significantly increased Bcl-2 expression,significantly decreased Beclin-1 and LC3-Ⅱ protein expressions and number of TUNEL postive neurons,and statistically increased p-Akt and p-mTOR levels in the TBI+COG1410 (2 mg) group were noted (P＜0.05).Conclusion ApoE peptide is effective in reducing the excessive neuronal apoptosis and neurological dysfunctions caused by excessive neuronal autophagy after TBI,which is associated with the modulation of Akt/mTOR related pathway.